Difference between revisions of "Team:Kyoto/Notebook"

 
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{{Kyoto}}
 
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{{Kyoto/header}}
 
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   .keln_researcher{
 
   .keln_researcher{
 
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p.keln_note{
 
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}   
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#jump{
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</style>
 
</style>
  
 
<body>
 
<body>
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  <div id="jump"><a href="#kyoto-calender"><img src="https://static.igem.org/mediawiki/2017/c/c5/Kyoto_notebook_jump.png"></a></div>
 
   <div id="BACKGROUND">
 
   <div id="BACKGROUND">
 
   <div id="wrapper">
 
   <div id="wrapper">
−
   <header>
+
    
−
    <ul class="nv">
+
−
        <li><a href="https://2017.igem.org/Team:Kyoto">Home</a></li>
+
−
        <li><a href="https://2017.igem.org/team:Kyoto/Project">Project</a>
+
−
            <ul>
+
−
              <li><a href="https://2017.igem.org/team:Kyoto/Project">Description</a></li>
+
−
              <li><a href="https://2017.igem.org/Team:Kyoto/Results">Results</a></li>
+
−
              <li><a href="https://2017.igem.org/Team:Kyoto/Demonstrate">Demonstrate</li>
+
−
              <li><a href="https://2017.igem.org/Team:Kyoto/Basic_Part">Basic parts</a></li>
+
−
              <li><a href="https://2017.igem.org/Team:Kyoto/Composite_Part">Composite parts</a></li>
+
−
            </ul>
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−
        </li><li><a href="https://2017.igem.org/Team:Kyoto/Experiments">Material&amp;Methods</a></li><li><a href="https://2017.igem.org/Team:Kyoto/Achievements">Medal criteria</a>
+
−
            <ul>
+
−
              <li><a href="https://2017.igem.org/Team:Kyoto/Achievements">Achievements</a></li>
+
−
              <li><a href="https://2017.igem.org/Team:Kyoto/HP/Silver">HP/Silver</a></li>
+
−
              <li><a href="https://2017.igem.org/Team:Kyoto/HP/Gold_Integrated">HP/Gold</a></li>
+
−
            </ul></li><li><a href="https://2017.igem.org/Team:Kyoto/Notebook">Notebook</a></li><li><a href="https://2017.igem.org/Team:Kyoto/integrated">Practice</a>
+
−
            <ul>
+
−
              <li><a href="https://2017.igem.org/Team:Kyoto/integrated">Human Practice</li>
+
−
              <li><a href="https://2017.igem.org/Team:Kyoto/integrated2">Integrated Human Practice</li>
+
−
              <li><a href="https://2017.igem.org/Team:Kyoto/InterLab">Public Engagement</li>
+
−
              <li><a href="https://2017.igem.org/Team:Kyoto/Collaborations">Collaboration</li>
+
−
            </ul></li><li><a href="https://2017.igem.org/Team:Kyoto/Team">Team</a>
+
−
            <ul>
+
−
              <li><a href="https://2017.igem.org/Team:Kyoto/Team">Members</li>
+
−
              <li><a href="https://2017.igem.org/Team:Kyoto/Attributions">Attributions</li>
+
−
            </ul></li><li><a href="https://2017.igem.org/Team:Kyoto/Safety">Safety</a></li>
+
−
      </ul>
+
−
  </header>
+
 
<h1>Notebook</h1>
 
<h1>Notebook</h1>
  
  
−
<div class="kyoto-calendar">
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<div class="kyoto-calendar" id="kyoto-calender">
  
  
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         <td><a href="#0921">21</a></td>
 
         <td><a href="#0921">21</a></td>
 
         <td><a href="#0922">22</a></td>
 
         <td><a href="#0922">22</a></td>
−
         <td><a href="#0923">23</a></td>
+
         <td>23</td>
 
         <td>24</td>
 
         <td>24</td>
 
         <td>25</td>
 
         <td>25</td>
−
         <td><a href="#0926">26</a></td>
+
         <td>26</td>
 
       </tr>
 
       </tr>
 
       <tr>
 
       <tr>
−
         <td><a href="#0927">27</a></td>
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         <td>27</td>
 
         <td>28</td>
 
         <td>28</td>
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         <td><a href="#0929">29</a></td>
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         <td>29</td>
−
         <td><a href="#0930">30</a></td>
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         <td>30</td>
 
         <td>31</td>
 
         <td>31</td>
 
         <td></td>
 
         <td></td>
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         <td></td>
 
         <td></td>
 
         <td></td>
 
         <td></td>
−
         <td><a href="#1001">1</a></td>
+
         <td>1</td>
 
         <td>2</td>
 
         <td>2</td>
 
         <td>3</td>
 
         <td>3</td>
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         <td><a href="#1007">7</a></td>
 
         <td><a href="#1007">7</a></td>
 
         <td>8</td>
 
         <td>8</td>
−
         <td>9</td>
+
         <td><a href="#1009">9</a></td>
 
         <td>10</td>
 
         <td>10</td>
−
         <td><a href="#1011">11</a></td>
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         <td>11</td>
 
         <td><a href="#1012">12</a></td>
 
         <td><a href="#1012">12</a></td>
 
       </tr>
 
       </tr>
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         <td><a href="#1014">14</a></td>
 
         <td><a href="#1014">14</a></td>
 
         <td><a href="#1015">15</a></td>
 
         <td><a href="#1015">15</a></td>
−
         <td><a href="#1016">16</a></td>
+
         <td>16</td>
 
         <td><a href="#1017">17</a></td>
 
         <td><a href="#1017">17</a></td>
 
         <td><a href="#1018">18</a></td>
 
         <td><a href="#1018">18</a></td>
−
         <td>19</td>
+
         <td><a href="#1019">19</a></td>
 
       </tr>
 
       </tr>
 
       <tr>
 
       <tr>
−
         <td>20</td>
+
         <td><a href="#1020">20</a></td>
 
         <td>21</td>
 
         <td>21</td>
−
         <td>22</td>
+
         <td><a href="#1022">22</a></td>
 
         <td>23</td>
 
         <td>23</td>
 
         <td>24</td>
 
         <td>24</td>
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<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>IK121</td><td>IK24</td><td>prt3</td></tr><tr><td>2</td><td>IK121</td><td>IK24</td><td>num99</td></tr><tr><td>3</td><td>IK121</td><td>IK24</td><td>OR1-1</td></tr><tr><td>4</td><td>IK121</td><td>IK24</td><td>OR1-2</td></tr><tr><td>5</td><td>IK121</td><td>IK24</td><td>OR1-3</td></tr><tr><td>6</td><td>IK121</td><td>IK24</td><td>OR1-5</td></tr><tr><td>7</td><td>IK121</td><td>IK24</td><td>OR1-4</td></tr>
 
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>IK121</td><td>IK24</td><td>prt3</td></tr><tr><td>2</td><td>IK121</td><td>IK24</td><td>num99</td></tr><tr><td>3</td><td>IK121</td><td>IK24</td><td>OR1-1</td></tr><tr><td>4</td><td>IK121</td><td>IK24</td><td>OR1-2</td></tr><tr><td>5</td><td>IK121</td><td>IK24</td><td>OR1-3</td></tr><tr><td>6</td><td>IK121</td><td>IK24</td><td>OR1-5</td></tr><tr><td>7</td><td>IK121</td><td>IK24</td><td>OR1-4</td></tr>
 
</table>
 
</table>
−
<p class="keln_note">PCR Polymerase : prime star max <br>98.0&deg;C : 3min<br>98.0&deg;C : 10sec<br>55.0&deg;C : 5sec<br>72.0&deg;C : 10sec<br>72.0&deg;C : 2min<br>4.0&deg;C : Infinate<br>X35 cycle</p><img src="https://static.igem.org/mediawiki/2017/b/be/IGKU_b2_150_0.jpeg">
+
<p class="keln_note">PCR Polymerase : prime star max <br>98.0&deg;C : 3min<br>98.0&deg;C : 10sec<br>55.0&deg;C : 5sec<br>72.0&deg;C : 10sec<br>72.0&deg;C : 2min<br>4.0&deg;C : Infinate<br>X35 cycle<img src="https://static.igem.org/mediawiki/2017/b/be/IGKU_b2_150_0.jpeg"></p><img src="https://static.igem.org/mediawiki/2017/b/be/IGKU_b2_150_0.jpeg">
 
<!-- IGKU_b2_150_1.jpeg -->
 
<!-- IGKU_b2_150_1.jpeg -->
 
<!-- IGKU_b2_150_2.jpeg -->
 
<!-- IGKU_b2_150_2.jpeg -->
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<!-- ############################################################## Book3 Tables Start ################################################## -->
 
<!-- ############################################################## Book3 Tables Start ################################################## -->
−
<!-- book3_tables.txt -->
+
 
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1004" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/04</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme digestion</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table"><tr><th>lane</th><th>Sample DNA name</th><th>DNA amount (ul)</th><th>RE1 name</th><th>RE1 amount(ul)</th><th>RE2 name</th><th>RE2 amount(ul)</th><th>Buffer name</th><th>Buffer amount(ul)</th><th>Total volume(ul)</th></tr><tr><td>1</td><td>Template 5</td><td>5.0</td><td>Xba</td><td>0.025</td><td>EcoR1</td><td>0.025</td><td>M</td><td>1.0</td><td>10.0</td></tr><tr><td>2</td><td>Template 5</td><td>5.0</td><td>Not1</td><td>0.025</td><td>EcoR1</td><td>0.025</td><td>H</td><td>1.0</td><td>10.0</td></tr><tr><td>3</td><td>Template 5</td><td>5.0</td><td>Not1</td><td>0.025</td><td>Xba1</td><td>0.025</td><td>H</td><td>1.0</td><td>10.0</td></tr>
 +
</table>
 +
<!-- IGKU_b3_008_0.jpeg -->
 +
<!-- IGKU_b3_008_1.jpeg -->
 +
<!-- IGKU_b3_008_2.jpeg -->
 +
<!-- IGKU_b3_008_3.jpeg -->
 +
<!-- IGKU_b3_008_4.jpeg -->
 +
<!-- IGKU_b3_008_5.jpeg -->
 +
<!-- IGKU_b3_008_6.jpeg -->
 +
<!-- IGKU_b3_008_7.jpeg -->
 +
<!-- IGKU_b3_008_8.jpeg -->
 +
<!-- IGKU_b3_008_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1004" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/04</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">ohara</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>OD21</td><td>OD23</td><td>eGFP plasmid</td></tr><tr><td>2</td><td>OD21</td><td>OD23</td><td>eGFP plasmid</td></tr><tr><td>3</td><td>OD22</td><td>OD23</td><td>eGFP plasmid</td></tr><tr><td>4</td><td>OD22</td><td>OD23</td><td>eGFP plasmid</td></tr><tr><td>5</td><td>OD31</td><td>OD32</td><td>T7-RRE c</td></tr><tr><td>6</td><td>OD31</td><td>OD32</td><td>T7-RRE c</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : prime star max <br>98.0&deg;C : 10sec<br>98.0&deg;C : 10sec<br>55.0&deg;C : 5sec<br>72.0&deg;C : 10sec<br>72.0&deg;C : 10sec<br>15.0&deg;C : Infinate<br>X35 cycle</p>
 +
<!-- IGKU_b3_009_0.jpeg -->
 +
<!-- IGKU_b3_009_1.jpeg -->
 +
<!-- IGKU_b3_009_2.jpeg -->
 +
<!-- IGKU_b3_009_3.jpeg -->
 +
<!-- IGKU_b3_009_4.jpeg -->
 +
<!-- IGKU_b3_009_5.jpeg -->
 +
<!-- IGKU_b3_009_6.jpeg -->
 +
<!-- IGKU_b3_009_7.jpeg -->
 +
<!-- IGKU_b3_009_8.jpeg -->
 +
<!-- IGKU_b3_009_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1004" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/04</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme digestion</h4></span>
 +
<span class="keln_researcher">Yoshimoto</span>
 +
<table class = "keln_table"><tr><th>lane</th><th>Sample DNA name</th><th>DNA amount (ul)</th><th>RE1 name</th><th>RE1 amount(ul)</th><th>RE2 name</th><th>RE2 amount(ul)</th><th>Buffer name</th><th>Buffer amount(ul)</th><th>Total volume(ul)</th></tr><tr><td>1</td><td>pSB1C3</td><td>15.0</td><td>Xba1</td><td>5.0</td><td>Spe1</td><td>5.0</td><td>M</td><td>12.0</td><td>100.0</td></tr><tr><td>2</td><td>pSB1C3</td><td>15.0</td><td>Xba1</td><td>5.0</td><td>Pst1</td><td>5.0</td><td>M</td><td>12.0</td><td>100.0</td></tr>
 +
</table>
 +
<!-- IGKU_b3_033_0.jpeg -->
 +
<!-- IGKU_b3_033_1.jpeg -->
 +
<!-- IGKU_b3_033_2.jpeg -->
 +
<!-- IGKU_b3_033_3.jpeg -->
 +
<!-- IGKU_b3_033_4.jpeg -->
 +
<!-- IGKU_b3_033_5.jpeg -->
 +
<!-- IGKU_b3_033_6.jpeg -->
 +
<!-- IGKU_b3_033_7.jpeg -->
 +
<!-- IGKU_b3_033_8.jpeg -->
 +
<!-- IGKU_b3_033_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1005" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/05</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>vf </td><td>vr</td><td>Colony1~20(191)</td></tr><tr><td>2</td><td>vf </td><td>vr</td><td>Colony21~40(192)</td></tr><tr><td>3</td><td>vf </td><td>vr</td><td>Colony41~60(193)</td></tr><tr><td>4</td><td>vf </td><td>vr</td><td>Colony61~80(194)</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : prime star max <br>98.0&deg;C : 10sec<br>98.0&deg;C : 10sec<br>55.0&deg;C : 5sec<br>72.0&deg;C : 10sec<br>72.0&deg;C : 10sec<br>15.0&deg;C : Infinate<br>X35 cycle</p>
 +
<!-- IGKU_b3_010_0.jpeg -->
 +
<!-- IGKU_b3_010_1.jpeg -->
 +
<!-- IGKU_b3_010_2.jpeg -->
 +
<!-- IGKU_b3_010_3.jpeg -->
 +
<!-- IGKU_b3_010_4.jpeg -->
 +
<!-- IGKU_b3_010_5.jpeg -->
 +
<!-- IGKU_b3_010_6.jpeg -->
 +
<!-- IGKU_b3_010_7.jpeg -->
 +
<!-- IGKU_b3_010_8.jpeg -->
 +
<!-- IGKU_b3_010_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1005" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/05</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme digestion</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table"><tr><th>lane</th><th>Sample DNA name</th><th>DNA amount (ul)</th><th>RE1 name</th><th>RE1 amount(ul)</th><th>Buffer name</th><th>Buffer amount(ul)</th><th>Total volume(ul)</th></tr><tr><td>1</td><td>template</td><td>5.0</td><td>Pst1</td><td>0.05</td><td>10xH</td><td>1.0</td><td>10.0</td></tr>
 +
</table>
 +
<!-- IGKU_b3_013_0.jpeg -->
 +
<!-- IGKU_b3_013_1.jpeg -->
 +
<!-- IGKU_b3_013_2.jpeg -->
 +
<!-- IGKU_b3_013_3.jpeg -->
 +
<!-- IGKU_b3_013_4.jpeg -->
 +
<!-- IGKU_b3_013_5.jpeg -->
 +
<!-- IGKU_b3_013_6.jpeg -->
 +
<!-- IGKU_b3_013_7.jpeg -->
 +
<!-- IGKU_b3_013_8.jpeg -->
 +
<!-- IGKU_b3_013_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1006" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/06</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>OD41</td><td>OD42</td><td>Gal-GFP</td></tr><tr><td>2</td><td>OD41</td><td>OD43</td><td>Gal-GFP</td></tr><tr><td>3</td><td>OD41</td><td>OD44</td><td>Gal-GFP</td></tr><tr><td>4</td><td>IK61</td><td>OD52</td><td>Gal-GFP</td></tr><tr><td>5</td><td>OD53</td><td>OD54</td><td>Gal-GFP</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : prime star max <br>98.0&deg;C : 10sec<br>98.0&deg;C : 10sec<br>55.0&deg;C : 5sec<br>72.0&deg;C : 10sec<br>72.0&deg;C : 10sec<br>15.0&deg;C : Infinate<br>X35 cycle</p>
 +
<!-- IGKU_b3_012_0.jpeg -->
 +
<!-- IGKU_b3_012_1.jpeg -->
 +
<!-- IGKU_b3_012_2.jpeg -->
 +
<!-- IGKU_b3_012_3.jpeg -->
 +
<!-- IGKU_b3_012_4.jpeg -->
 +
<!-- IGKU_b3_012_5.jpeg -->
 +
<!-- IGKU_b3_012_6.jpeg -->
 +
<!-- IGKU_b3_012_7.jpeg -->
 +
<!-- IGKU_b3_012_8.jpeg -->
 +
<!-- IGKU_b3_012_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1007" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/07</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>IK61</td><td>OD54</td><td>T7-RRE</td></tr><tr><td>2</td><td>IK61</td><td>OD54</td><td>fukuda GFP</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : pfu <br>95.0&deg;C : 2min<br>95.0&deg;C : 1min<br>55.0&deg;C : 30sec<br>72.0&deg;C : 4min<br>72.0&deg;C : 4min<br>15.0&deg;C : Infinate<br>X40 cycle</p>
 +
<!-- IGKU_b3_014_0.jpeg -->
 +
<!-- IGKU_b3_014_1.jpeg -->
 +
<!-- IGKU_b3_014_2.jpeg -->
 +
<!-- IGKU_b3_014_3.jpeg -->
 +
<!-- IGKU_b3_014_4.jpeg -->
 +
<!-- IGKU_b3_014_5.jpeg -->
 +
<!-- IGKU_b3_014_6.jpeg -->
 +
<!-- IGKU_b3_014_7.jpeg -->
 +
<!-- IGKU_b3_014_8.jpeg -->
 +
<!-- IGKU_b3_014_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1007" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/07</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>OD11</td><td>OD12</td><td>GPD-gfp</td></tr><tr><td>2</td><td>OD11</td><td>OD13</td><td>GPD-gfp</td></tr><tr><td>3</td><td>OD41</td><td>OD42</td><td>Gal-GFP</td></tr><tr><td>4</td><td>OD41</td><td>OD43</td><td>Gal-GFP</td></tr><tr><td>5</td><td>OD41</td><td>OD44</td><td>Gal-GFP</td></tr><tr><td>6</td><td>OD31</td><td>OD32</td><td>T7-RRE</td></tr><tr><td>7</td><td>IK61</td><td>IK54</td><td>fukuda GFP + T7RRE</td></tr><tr><td>8</td><td>IK61</td><td>IK54</td><td>fukuda GFP + T7RRE</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : pfu <br>95.0&deg;C : 2min<br>95.0&deg;C : 1min<br>55.0&deg;C : 30sec<br>72.0&deg;C : 4min<br>72.0&deg;C : 4min<br>15.0&deg;C : Infinate<br>X40 cycle</p>
 +
<!-- IGKU_b3_015_0.jpeg -->
 +
<!-- IGKU_b3_015_1.jpeg -->
 +
<!-- IGKU_b3_015_2.jpeg -->
 +
<!-- IGKU_b3_015_3.jpeg -->
 +
<!-- IGKU_b3_015_4.jpeg -->
 +
<!-- IGKU_b3_015_5.jpeg -->
 +
<!-- IGKU_b3_015_6.jpeg -->
 +
<!-- IGKU_b3_015_7.jpeg -->
 +
<!-- IGKU_b3_015_8.jpeg -->
 +
<!-- IGKU_b3_015_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1009" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/09</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme digestion</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table"><tr><th>lane</th><th>Sample DNA name</th><th>DNA amount (ul)</th><th>RE1 name</th><th>RE1 amount(ul)</th><th>RE2 name</th><th>RE2 amount(ul)</th><th>Buffer name</th><th>Buffer amount(ul)</th><th>Total volume(ul)</th></tr><tr><td>1</td><td>T7 RRE</td><td>15.0</td><td>Xba1</td><td>3.0</td><td>Pst1</td><td>3.0</td><td>M</td><td>3.0</td><td>30.0</td></tr>
 +
</table>
 +
<!-- IGKU_b3_019_0.jpeg -->
 +
<!-- IGKU_b3_019_1.jpeg -->
 +
<!-- IGKU_b3_019_2.jpeg -->
 +
<!-- IGKU_b3_019_3.jpeg -->
 +
<!-- IGKU_b3_019_4.jpeg -->
 +
<!-- IGKU_b3_019_5.jpeg -->
 +
<!-- IGKU_b3_019_6.jpeg -->
 +
<!-- IGKU_b3_019_7.jpeg -->
 +
<!-- IGKU_b3_019_8.jpeg -->
 +
<!-- IGKU_b3_019_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1009" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/09</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>IK61</td><td>IK54</td><td>GFP+RRE</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : pfu <br>95.0&deg;C : 2min<br>95.0&deg;C : 1min<br>55.0&deg;C : 30sec<br>72.0&deg;C : 4min<br>72.0&deg;C : 4min<br>15.0&deg;C : Infinate<br>X40 cycle</p>
 +
<!-- IGKU_b3_020_0.jpeg -->
 +
<!-- IGKU_b3_020_1.jpeg -->
 +
<!-- IGKU_b3_020_2.jpeg -->
 +
<!-- IGKU_b3_020_3.jpeg -->
 +
<!-- IGKU_b3_020_4.jpeg -->
 +
<!-- IGKU_b3_020_5.jpeg -->
 +
<!-- IGKU_b3_020_6.jpeg -->
 +
<!-- IGKU_b3_020_7.jpeg -->
 +
<!-- IGKU_b3_020_8.jpeg -->
 +
<!-- IGKU_b3_020_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1009" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/09</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>OD41</td><td>OD42</td><td>Gal-gfp</td></tr><tr><td>2</td><td>OD41</td><td>OD44</td><td>Gal-gfp</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : pfu <br>95.0&deg;C : 2min<br>95.0&deg;C : 1min<br>55.0&deg;C : 30sec<br>72.0&deg;C : 4min<br>72.0&deg;C : 4min<br>15.0&deg;C : Infinate<br>X35 cycle</p>
 +
<!-- IGKU_b3_021_0.jpeg -->
 +
<!-- IGKU_b3_021_1.jpeg -->
 +
<!-- IGKU_b3_021_2.jpeg -->
 +
<!-- IGKU_b3_021_3.jpeg -->
 +
<!-- IGKU_b3_021_4.jpeg -->
 +
<!-- IGKU_b3_021_5.jpeg -->
 +
<!-- IGKU_b3_021_6.jpeg -->
 +
<!-- IGKU_b3_021_7.jpeg -->
 +
<!-- IGKU_b3_021_8.jpeg -->
 +
<!-- IGKU_b3_021_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1012" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/12</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>OD61</td><td>OD12</td><td>TDH3-eGFP</td></tr><tr><td>2</td><td>OD62</td><td>OD12</td><td>TDH3-eGFP</td></tr><tr><td>3</td><td>OD63</td><td>OD12</td><td>TDH3-eGFP</td></tr><tr><td>4</td><td>OD61</td><td>OD13</td><td>TDH3-eGFP</td></tr><tr><td>5</td><td>OD62</td><td>OD13</td><td>TDH3-eGFP</td></tr><tr><td>6</td><td>OD63</td><td>OD13</td><td>TDH3-eGFP</td></tr><tr><td>7</td><td>OD73</td><td>IK62</td><td>GFP</td></tr><tr><td>8</td><td>OD45</td><td>OD42</td><td>GFP</td></tr><tr><td>9</td><td>OD45</td><td>OD43</td><td>GFP</td></tr><tr><td>10</td><td>OD45</td><td>OD44</td><td>GFP</td></tr><tr><td>11</td><td>IK61</td><td>OD74</td><td>GFP</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : KAPA HiFi fast <br>95.0&deg;C : 5min<br>98.0&deg;C : 20sec<br>58.0&deg;C : 15sec<br>72.0&deg;C : 40sec<br>72.0&deg;C : 5min<br>4.0&deg;C : Infinate<br>X35 cycle</p>
 +
<!-- IGKU_b3_025_0.jpeg -->
 +
<!-- IGKU_b3_025_1.jpeg -->
 +
<!-- IGKU_b3_025_2.jpeg -->
 +
<!-- IGKU_b3_025_3.jpeg -->
 +
<!-- IGKU_b3_025_4.jpeg -->
 +
<!-- IGKU_b3_025_5.jpeg -->
 +
<!-- IGKU_b3_025_6.jpeg -->
 +
<!-- IGKU_b3_025_7.jpeg -->
 +
<!-- IGKU_b3_025_8.jpeg -->
 +
<!-- IGKU_b3_025_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1012" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/12</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme digestion</h4></span>
 +
<span class="keln_researcher">Yoshimoto</span>
 +
<table class = "keln_table"><tr><th>lane</th><th>Sample DNA name</th><th>DNA amount (ul)</th><th>RE1 name</th><th>RE1 amount(ul)</th><th>Buffer name</th><th>Buffer amount(ul)</th><th>Total volume(ul)</th></tr><tr><td>1</td><td>Template14</td><td>5.0</td><td>Xba1</td><td>1.0</td><td>M</td><td>1.0</td><td>10.0</td></tr><tr><td>2</td><td>Template100</td><td>5.0</td><td>Xba1</td><td>1.0</td><td>M</td><td>1.0</td><td>10.0</td></tr>
 +
</table>
 +
<!-- IGKU_b3_027_0.jpeg -->
 +
<!-- IGKU_b3_027_1.jpeg -->
 +
<!-- IGKU_b3_027_2.jpeg -->
 +
<!-- IGKU_b3_027_3.jpeg -->
 +
<!-- IGKU_b3_027_4.jpeg -->
 +
<!-- IGKU_b3_027_5.jpeg -->
 +
<!-- IGKU_b3_027_6.jpeg -->
 +
<!-- IGKU_b3_027_7.jpeg -->
 +
<!-- IGKU_b3_027_8.jpeg -->
 +
<!-- IGKU_b3_027_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1012" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/12</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>OD62</td><td>OD12</td><td>yeast genome</td></tr><tr><td>2</td><td>OD62</td><td>OD13</td><td>yeast genome</td></tr><tr><td>3</td><td>OD73</td><td>IK62</td><td>fukuda gfp</td></tr><tr><td>4</td><td>OD45</td><td>OD42</td><td>gal gfp</td></tr><tr><td>5</td><td>IK61</td><td>OD74</td><td>fukuda gfp</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : KAPA hifi  <br>95.0&deg;C : 5min<br>98.0&deg;C : 20sec<br>58.0&deg;C : 15sec<br>72.0&deg;C : 40sec<br>72.0&deg;C : 5min<br>4.0&deg;C : Infinate<br>X35 cycle</p>
 +
<!-- IGKU_b3_028_0.jpeg -->
 +
<!-- IGKU_b3_028_1.jpeg -->
 +
<!-- IGKU_b3_028_2.jpeg -->
 +
<!-- IGKU_b3_028_3.jpeg -->
 +
<!-- IGKU_b3_028_4.jpeg -->
 +
<!-- IGKU_b3_028_5.jpeg -->
 +
<!-- IGKU_b3_028_6.jpeg -->
 +
<!-- IGKU_b3_028_7.jpeg -->
 +
<!-- IGKU_b3_028_8.jpeg -->
 +
<!-- IGKU_b3_028_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1013" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/13</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>OD62</td><td>OD12</td><td>yeast genome</td></tr><tr><td>2</td><td>OD62</td><td>OD12</td><td>yeast genome</td></tr><tr><td>3</td><td>OD62</td><td>OD13</td><td>TDH3-eGFP</td></tr><tr><td>4</td><td>OD62</td><td>OD13</td><td>TDH3-eGFP</td></tr><tr><td>5</td><td>OD73</td><td>IK62</td><td>fukuda GFP</td></tr><tr><td>6</td><td>IK61</td><td>OD71</td><td>fukuda GFP</td></tr><tr><td>7</td><td>OD72</td><td>OD54</td><td>T7-RRE</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : KAPA hifi <br>95.0&deg;C : 5min<br>98.0&deg;C : 20sec<br>58.0&deg;C : 15sec<br>72.0&deg;C : 40sec<br>72.0&deg;C : 5min<br>4.0&deg;C : Infinate<br>X35 cycle</p>
 +
<!-- IGKU_b3_029_0.jpeg -->
 +
<!-- IGKU_b3_029_1.jpeg -->
 +
<!-- IGKU_b3_029_2.jpeg -->
 +
<!-- IGKU_b3_029_3.jpeg -->
 +
<!-- IGKU_b3_029_4.jpeg -->
 +
<!-- IGKU_b3_029_5.jpeg -->
 +
<!-- IGKU_b3_029_6.jpeg -->
 +
<!-- IGKU_b3_029_7.jpeg -->
 +
<!-- IGKU_b3_029_8.jpeg -->
 +
<!-- IGKU_b3_029_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1014" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/14</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>OD62</td><td>OD12</td><td>yeast genome</td></tr><tr><td>2</td><td>OD62</td><td>OD13</td><td>yeast genome</td></tr><tr><td>3</td><td>OD45</td><td>OD42</td><td>gal1 GFP</td></tr><tr><td>4</td><td>IK61</td><td>OD74</td><td>fukuda GFP</td></tr><tr><td>5</td><td>OD73</td><td>IK62</td><td>fukuda GFP</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : KAPA <br>95.0&deg;C : 5min<br>98.0&deg;C : 20sec<br>58.0&deg;C : 15sec<br>72.0&deg;C : 40sec<br>72.0&deg;C : 5min<br>4.0&deg;C : Infinate<br>X35 cycle</p>
 +
<!-- IGKU_b3_032_0.jpeg -->
 +
<!-- IGKU_b3_032_1.jpeg -->
 +
<!-- IGKU_b3_032_2.jpeg -->
 +
<!-- IGKU_b3_032_3.jpeg -->
 +
<!-- IGKU_b3_032_4.jpeg -->
 +
<!-- IGKU_b3_032_5.jpeg -->
 +
<!-- IGKU_b3_032_6.jpeg -->
 +
<!-- IGKU_b3_032_7.jpeg -->
 +
<!-- IGKU_b3_032_8.jpeg -->
 +
<!-- IGKU_b3_032_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: yoshimoto -->
 +
<div class="keln_container">
 +
<a name="1014" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/14</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">ohara</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>IK103</td><td>IK104</td><td>165 gBlock</td></tr><tr><td>2</td><td>IK107</td><td>IK108</td><td>161 gBlock</td></tr><tr><td>3</td><td>IK101</td><td>IK102</td><td>161 gBlock</td></tr><tr><td>4</td><td>IK105</td><td>IK106</td><td>163 gBlock</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : yoshimoto <br>95.0&deg;C : 5min<br>98.0&deg;C : 20sec<br>58.0&deg;C : 15sec<br>72.0&deg;C : 40sec<br>72.0&deg;C : 5min<br>4.0&deg;C : Infinate<br>X35 cycle</p>
 +
<!-- IGKU_b3_034_0.jpeg -->
 +
<!-- IGKU_b3_034_1.jpeg -->
 +
<!-- IGKU_b3_034_2.jpeg -->
 +
<!-- IGKU_b3_034_3.jpeg -->
 +
<!-- IGKU_b3_034_4.jpeg -->
 +
<!-- IGKU_b3_034_5.jpeg -->
 +
<!-- IGKU_b3_034_6.jpeg -->
 +
<!-- IGKU_b3_034_7.jpeg -->
 +
<!-- IGKU_b3_034_8.jpeg -->
 +
<!-- IGKU_b3_034_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1014" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/14</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>OD45</td><td>OD42</td><td>gal1 GFP</td></tr><tr><td>2</td><td>IK61</td><td>OD74</td><td>fukuda GFP</td></tr><tr><td>3</td><td>OD73</td><td>IK62</td><td>fukuda GFP</td></tr><tr><td>4</td><td>IK61</td><td>OD54</td><td>fukuda GFP</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : KAPA hifi <br>95.0&deg;C : 5min<br>98.0&deg;C : 20sec<br>58.0&deg;C : 15sec<br>72.0&deg;C : 40sec<br>72.0&deg;C : 5min<br>4.0&deg;C : Infinate<br>X35 cycle</p>
 +
<!-- IGKU_b3_035_0.jpeg -->
 +
<!-- IGKU_b3_035_1.jpeg -->
 +
<!-- IGKU_b3_035_2.jpeg -->
 +
<!-- IGKU_b3_035_3.jpeg -->
 +
<!-- IGKU_b3_035_4.jpeg -->
 +
<!-- IGKU_b3_035_5.jpeg -->
 +
<!-- IGKU_b3_035_6.jpeg -->
 +
<!-- IGKU_b3_035_7.jpeg -->
 +
<!-- IGKU_b3_035_8.jpeg -->
 +
<!-- IGKU_b3_035_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1014" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/14</h3></span>
 +
<span class="keln_exp"><h4>Restriction Enzyme digestion</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table"><tr><th>lane</th><th>Sample DNA name</th><th>DNA amount (ul)</th><th>RE1 name</th><th>RE1 amount(ul)</th><th>Buffer name</th><th>Buffer amount(ul)</th><th>Total volume(ul)</th></tr><tr><td>1</td><td>template</td><td>15.0</td><td>Xba1</td><td>3.0</td><td>M</td><td>3.0</td><td>100.0</td></tr><tr><td>2</td><td>template</td><td>15.0</td><td>Xba1</td><td>5.0</td><td>M</td><td>3.0</td><td>100.0</td></tr>
 +
</table>
 +
<!-- IGKU_b3_036_0.jpeg -->
 +
<!-- IGKU_b3_036_1.jpeg -->
 +
<!-- IGKU_b3_036_2.jpeg -->
 +
<!-- IGKU_b3_036_3.jpeg -->
 +
<!-- IGKU_b3_036_4.jpeg -->
 +
<!-- IGKU_b3_036_5.jpeg -->
 +
<!-- IGKU_b3_036_6.jpeg -->
 +
<!-- IGKU_b3_036_7.jpeg -->
 +
<!-- IGKU_b3_036_8.jpeg -->
 +
<!-- IGKU_b3_036_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1014" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/14</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>OD31</td><td>OD32</td><td>T7+RRE</td></tr><tr><td>2</td><td>OD22</td><td>OD23</td><td>eGFP</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : KAPA hifi <br>95.0&deg;C : 5min<br>98.0&deg;C : 20sec<br>58.0&deg;C : 15sec<br>72.0&deg;C : 40sec<br>72.0&deg;C : 5min<br>4.0&deg;C : Infinate<br>X35 cycle</p>
 +
<!-- IGKU_b3_037_0.jpeg -->
 +
<!-- IGKU_b3_037_1.jpeg -->
 +
<!-- IGKU_b3_037_2.jpeg -->
 +
<!-- IGKU_b3_037_3.jpeg -->
 +
<!-- IGKU_b3_037_4.jpeg -->
 +
<!-- IGKU_b3_037_5.jpeg -->
 +
<!-- IGKU_b3_037_6.jpeg -->
 +
<!-- IGKU_b3_037_7.jpeg -->
 +
<!-- IGKU_b3_037_8.jpeg -->
 +
<!-- IGKU_b3_037_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1015" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/15</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>IK61</td><td>OD54</td><td>6+7</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : KAPA hifi <br>95.0&deg;C : 5min<br>98.0&deg;C : 20sec<br>58.0&deg;C : 15sec<br>72.0&deg;C : 40sec<br>72.0&deg;C : 5min<br>4.0&deg;C : Infinate<br>X35 cycle</p>
 +
<!-- IGKU_b3_038_0.jpeg -->
 +
<!-- IGKU_b3_038_1.jpeg -->
 +
<!-- IGKU_b3_038_2.jpeg -->
 +
<!-- IGKU_b3_038_3.jpeg -->
 +
<!-- IGKU_b3_038_4.jpeg -->
 +
<!-- IGKU_b3_038_5.jpeg -->
 +
<!-- IGKU_b3_038_6.jpeg -->
 +
<!-- IGKU_b3_038_7.jpeg -->
 +
<!-- IGKU_b3_038_8.jpeg -->
 +
<!-- IGKU_b3_038_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1015" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/15</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>IK85</td><td>IK86</td><td>rev</td></tr><tr><td>2</td><td>IK87</td><td>Ik88</td><td>rev</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : KAPA hifi <br>95.0&deg;C : 5min<br>98.0&deg;C : 20sec<br>58.0&deg;C : 15sec<br>72.0&deg;C : 40sec<br>72.0&deg;C : 5min<br>4.0&deg;C : Infinate<br>X35 cycle<br>PCR pulification: 1~2<br>Gel extraction: 1~2</p>
 +
<!-- IGKU_b3_039_0.jpeg -->
 +
<!-- IGKU_b3_039_1.jpeg -->
 +
<!-- IGKU_b3_039_2.jpeg -->
 +
<!-- IGKU_b3_039_3.jpeg -->
 +
<!-- IGKU_b3_039_4.jpeg -->
 +
<!-- IGKU_b3_039_5.jpeg -->
 +
<!-- IGKU_b3_039_6.jpeg -->
 +
<!-- IGKU_b3_039_7.jpeg -->
 +
<!-- IGKU_b3_039_8.jpeg -->
 +
<!-- IGKU_b3_039_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1015" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/15</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>OD31</td><td>OD32</td><td>T7+RRE</td></tr><tr><td>2</td><td>OD22</td><td>OD23</td><td>TDH3promoter</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : KAPA hifi <br>95.0&deg;C : 5min<br>98.0&deg;C : 20sec<br>58.0&deg;C : 15sec<br>72.0&deg;C : 40sec<br>72.0&deg;C : 5min<br>4.0&deg;C : Infinate<br>X35 cycle</p>
 +
<!-- IGKU_b3_040_0.jpeg -->
 +
<!-- IGKU_b3_040_1.jpeg -->
 +
<!-- IGKU_b3_040_2.jpeg -->
 +
<!-- IGKU_b3_040_3.jpeg -->
 +
<!-- IGKU_b3_040_4.jpeg -->
 +
<!-- IGKU_b3_040_5.jpeg -->
 +
<!-- IGKU_b3_040_6.jpeg -->
 +
<!-- IGKU_b3_040_7.jpeg -->
 +
<!-- IGKU_b3_040_8.jpeg -->
 +
<!-- IGKU_b3_040_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1015" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/15</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>OD31</td><td>OD32</td><td>T7+RRE</td></tr><tr><td>2</td><td>OD22</td><td>OD23</td><td>TDH3promoter</td></tr><tr><td>3</td><td>OD45</td><td>OD42</td><td>Gal eGFP</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : KAPA hifi <br>95.0&deg;C : 5min<br>98.0&deg;C : 20sec<br>58.0&deg;C : 15sec<br>72.0&deg;C : 40sec<br>72.0&deg;C : 5min<br>4.0&deg;C : Infinate<br>X30 cycle</p>
 +
<!-- IGKU_b3_042_0.jpeg -->
 +
<!-- IGKU_b3_042_1.jpeg -->
 +
<!-- IGKU_b3_042_2.jpeg -->
 +
<!-- IGKU_b3_042_3.jpeg -->
 +
<!-- IGKU_b3_042_4.jpeg -->
 +
<!-- IGKU_b3_042_5.jpeg -->
 +
<!-- IGKU_b3_042_6.jpeg -->
 +
<!-- IGKU_b3_042_7.jpeg -->
 +
<!-- IGKU_b3_042_8.jpeg -->
 +
<!-- IGKU_b3_042_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1015" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/15</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>IK105</td><td>Ik106</td><td>Gblock 163</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : KAPA hifi <br>95.0&deg;C : 5min<br>98.0&deg;C : 20sec<br>58.0&deg;C : 15sec<br>72.0&deg;C : 40sec<br>72.0&deg;C : 5min<br>4.0&deg;C : Infinate<br>X35 cycle</p>
 +
<!-- IGKU_b3_043_0.jpeg -->
 +
<!-- IGKU_b3_043_1.jpeg -->
 +
<!-- IGKU_b3_043_2.jpeg -->
 +
<!-- IGKU_b3_043_3.jpeg -->
 +
<!-- IGKU_b3_043_4.jpeg -->
 +
<!-- IGKU_b3_043_5.jpeg -->
 +
<!-- IGKU_b3_043_6.jpeg -->
 +
<!-- IGKU_b3_043_7.jpeg -->
 +
<!-- IGKU_b3_043_8.jpeg -->
 +
<!-- IGKU_b3_043_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1015" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/15</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>IK105</td><td>IK108</td><td>Gal1 loop</td></tr><tr><td>2</td><td>IK101</td><td>Ik104</td><td>GFP loop</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : KAPA hifi <br>95.0&deg;C : 5min<br>98.0&deg;C : 20sec<br>58.0&deg;C : 15sec<br>72.0&deg;C : 40sec<br>72.0&deg;C : 5min<br>4.0&deg;C : Infinate<br>X35 cycle</p>
 +
<!-- IGKU_b3_044_0.jpeg -->
 +
<!-- IGKU_b3_044_1.jpeg -->
 +
<!-- IGKU_b3_044_2.jpeg -->
 +
<!-- IGKU_b3_044_3.jpeg -->
 +
<!-- IGKU_b3_044_4.jpeg -->
 +
<!-- IGKU_b3_044_5.jpeg -->
 +
<!-- IGKU_b3_044_6.jpeg -->
 +
<!-- IGKU_b3_044_7.jpeg -->
 +
<!-- IGKU_b3_044_8.jpeg -->
 +
<!-- IGKU_b3_044_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1015" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/15</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>IK85</td><td>IK86</td><td>Rev parts</td></tr><tr><td>2</td><td>IK87</td><td>IK88</td><td>Rev parts</td></tr><tr><td>3</td><td>IK61</td><td>OD54</td><td>T7-RRE-GFP</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : KAPA hifi <br>95.0&deg;C : 5min<br>98.0&deg;C : 20sec<br>58.0&deg;C : 15sec<br>72.0&deg;C : 40sec<br>72.0&deg;C : 5min<br>4.0&deg;C : Infinate<br>X30 cycle</p>
 +
<!-- IGKU_b3_045_0.jpeg -->
 +
<!-- IGKU_b3_045_1.jpeg -->
 +
<!-- IGKU_b3_045_2.jpeg -->
 +
<!-- IGKU_b3_045_3.jpeg -->
 +
<!-- IGKU_b3_045_4.jpeg -->
 +
<!-- IGKU_b3_045_5.jpeg -->
 +
<!-- IGKU_b3_045_6.jpeg -->
 +
<!-- IGKU_b3_045_7.jpeg -->
 +
<!-- IGKU_b3_045_8.jpeg -->
 +
<!-- IGKU_b3_045_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1017" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/17</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshitomo</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>IK61</td><td>OD54</td><td>4</td></tr><tr><td>2</td><td>IK61</td><td>OD54</td><td>5</td></tr><tr><td>3</td><td>IK61</td><td>OD54</td><td>7</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase : KAPA hifi <br>95.0&deg;C : 5min<br>98.0&deg;C : 20sec<br>58.0&deg;C : 15sec<br>72.0&deg;C : 40sec<br>72.0&deg;C : 5min<br>4.0&deg;C : Infinate<br>X30 cycle</p>
 +
<!-- IGKU_b3_048_0.jpeg -->
 +
<!-- IGKU_b3_048_1.jpeg -->
 +
<!-- IGKU_b3_048_2.jpeg -->
 +
<!-- IGKU_b3_048_3.jpeg -->
 +
<!-- IGKU_b3_048_4.jpeg -->
 +
<!-- IGKU_b3_048_5.jpeg -->
 +
<!-- IGKU_b3_048_6.jpeg -->
 +
<!-- IGKU_b3_048_7.jpeg -->
 +
<!-- IGKU_b3_048_8.jpeg -->
 +
<!-- IGKU_b3_048_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1017" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/17</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>IK87</td><td>IK88</td><td>Rev</td></tr><tr><td>2</td><td>IK105</td><td>IK108</td><td>Gal1-loop</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase :  <br>95.0&deg;C : 5min<br>98.0&deg;C : 20sec<br>55.0&deg;C : 15sec<br>72.0&deg;C : 40sec<br>72.0&deg;C : 5min<br>4.0&deg;C : Infinate<br>X30 cycle</p>
 +
<!-- IGKU_b3_049_0.jpeg -->
 +
<!-- IGKU_b3_049_1.jpeg -->
 +
<!-- IGKU_b3_049_2.jpeg -->
 +
<!-- IGKU_b3_049_3.jpeg -->
 +
<!-- IGKU_b3_049_4.jpeg -->
 +
<!-- IGKU_b3_049_5.jpeg -->
 +
<!-- IGKU_b3_049_6.jpeg -->
 +
<!-- IGKU_b3_049_7.jpeg -->
 +
<!-- IGKU_b3_049_8.jpeg -->
 +
<!-- IGKU_b3_049_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1018" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/18</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>IK105</td><td>IK108</td><td>template1</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase :  <br>95.0&deg;C : 5min<br>98.0&deg;C : 20sec<br>58.0&deg;C : 15sec<br>72.0&deg;C : 40sec<br>72.0&deg;C : 5min<br>4.0&deg;C : Infinate<br>X30 cycle</p>
 +
<!-- IGKU_b3_053_0.jpeg -->
 +
<!-- IGKU_b3_053_1.jpeg -->
 +
<!-- IGKU_b3_053_2.jpeg -->
 +
<!-- IGKU_b3_053_3.jpeg -->
 +
<!-- IGKU_b3_053_4.jpeg -->
 +
<!-- IGKU_b3_053_5.jpeg -->
 +
<!-- IGKU_b3_053_6.jpeg -->
 +
<!-- IGKU_b3_053_7.jpeg -->
 +
<!-- IGKU_b3_053_8.jpeg -->
 +
<!-- IGKU_b3_053_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1018" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/18</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>IK107</td><td>IK108</td><td>Gal1pro-loop</td></tr><tr><td>2</td><td>IK105</td><td>IK106</td><td>Gal1pro-loop</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase :  <br>95.0&deg;C : 5min<br>98.0&deg;C : 20sec<br>58.0&deg;C : 15sec<br>72.0&deg;C : 40sec<br>72.0&deg;C : 5min<br>4.0&deg;C : Infinate<br>X30 cycle</p>
 +
<!-- IGKU_b3_055_0.jpeg -->
 +
<!-- IGKU_b3_055_1.jpeg -->
 +
<!-- IGKU_b3_055_2.jpeg -->
 +
<!-- IGKU_b3_055_3.jpeg -->
 +
<!-- IGKU_b3_055_4.jpeg -->
 +
<!-- IGKU_b3_055_5.jpeg -->
 +
<!-- IGKU_b3_055_6.jpeg -->
 +
<!-- IGKU_b3_055_7.jpeg -->
 +
<!-- IGKU_b3_055_8.jpeg -->
 +
<!-- IGKU_b3_055_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1019" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/19</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>OD22</td><td>OD23</td><td>GPD-loop</td></tr><tr><td>2</td><td>IK87</td><td>IK88</td><td>Gal1-loop</td></tr><tr><td>3</td><td>IK105</td><td>IK108</td><td>T7-RRE</td></tr><tr><td>4</td><td>IK101</td><td>IK104</td><td>TDH3-eGFP</td></tr><tr><td>5</td><td>OD31</td><td>OD32</td><td>Rev</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase :  <br>95.0&deg;C : 5min<br>98.0&deg;C : 20sec<br>58.0&deg;C : 15sec<br>72.0&deg;C : 40sec<br>72.0&deg;C : 5min<br>4.0&deg;C : Infinate<br>X30 cycle</p>
 +
<!-- IGKU_b3_057_0.jpeg -->
 +
<!-- IGKU_b3_057_1.jpeg -->
 +
<!-- IGKU_b3_057_2.jpeg -->
 +
<!-- IGKU_b3_057_3.jpeg -->
 +
<!-- IGKU_b3_057_4.jpeg -->
 +
<!-- IGKU_b3_057_5.jpeg -->
 +
<!-- IGKU_b3_057_6.jpeg -->
 +
<!-- IGKU_b3_057_7.jpeg -->
 +
<!-- IGKU_b3_057_8.jpeg -->
 +
<!-- IGKU_b3_057_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1020" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/20</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>M13F</td><td>M13R</td><td>Colony 1-10 (RRE)</td></tr><tr><td>2</td><td>M13F</td><td>M13R</td><td>Colony11-17 (TDH3-eGFP)</td></tr><tr><td>3</td><td>M13F</td><td>M13R</td><td>Colony 18-21 (Rev)</td></tr><tr><td>4</td><td>M13F</td><td>M13R</td><td>Colony 22-23 (GPDp-loop)</td></tr><tr><td>5</td><td>M13F</td><td>M13R</td><td>Colony24-28 (GPDp-loop)</td></tr><tr><td>6</td><td>M13F</td><td>M13R</td><td>Colony 29-32 (T7-RRE)</td></tr><tr><td>7</td><td>M13F</td><td>M13R</td><td>Colony 33-35 (TDH3-eGFP)</td></tr><tr><td>8</td><td>M13F</td><td>M13R</td><td>Colony 36-37 (GPDp-loop)</td></tr><tr><td>9</td><td>M13F</td><td>M13R</td><td>Colony 38-40 (Gal1-loop)</td></tr><tr><td>10</td><td>M13F</td><td>M13R</td><td>Colony 41-43 (Rev)</td></tr><tr><td>11</td><td>M13F</td><td>M13R</td><td>Colony 44 (GPDp-loop)</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase :  <br>95.0&deg;C : 5min<br>98.0&deg;C : 20sec<br>58.0&deg;C : 15sec<br>72.0&deg;C : 40sec<br>72.0&deg;C : 5min<br>4.0&deg;C : Infinate<br>X30 cycle</p>
 +
<!-- IGKU_b3_058_0.jpeg -->
 +
<!-- IGKU_b3_058_1.jpeg -->
 +
<!-- IGKU_b3_058_2.jpeg -->
 +
<!-- IGKU_b3_058_3.jpeg -->
 +
<!-- IGKU_b3_058_4.jpeg -->
 +
<!-- IGKU_b3_058_5.jpeg -->
 +
<!-- IGKU_b3_058_6.jpeg -->
 +
<!-- IGKU_b3_058_7.jpeg -->
 +
<!-- IGKU_b3_058_8.jpeg -->
 +
<!-- IGKU_b3_058_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
<!-- Table Generated by KELN Author: ohara -->
 +
<div class="keln_container">
 +
<a name="1022" class = "kyoto-jump"></a>
 +
<span class="keln_date"><h3>10/22</h3></span>
 +
<span class="keln_exp"><h4>PCR</h4></span>
 +
<span class="keln_researcher">yoshimoto</span>
 +
<table class = "keln_table">
 +
<tr><th>lane</th><th>Primer Foward</th><th>Primer Revese</th><th>Template</th></tr><tr><td>1</td><td>OD22</td><td>OD23</td><td>TDH3 GFP</td></tr><tr><td>2</td><td>IK87</td><td>IK88</td><td>Rev</td></tr><tr><td>3</td><td>IK105</td><td>IK108</td><td>Gal1</td></tr><tr><td>4</td><td>IK101</td><td>IK104</td><td>GPD1</td></tr><tr><td>5</td><td>OD31</td><td>OD32</td><td>T7-RRE</td></tr>
 +
</table>
 +
<p class="keln_note">PCR Polymerase :  <br>95.0&deg;C : 5min<br>98.0&deg;C : 20sec<br>58.0&deg;C : 15sec<br>72.0&deg;C : 40sec<br>72.0&deg;C : 5min<br>4.0&deg;C : Infinate<br>X30 cycle</p>
 +
<!-- IGKU_b3_059_0.jpeg -->
 +
<!-- IGKU_b3_059_1.jpeg -->
 +
<!-- IGKU_b3_059_2.jpeg -->
 +
<!-- IGKU_b3_059_3.jpeg -->
 +
<!-- IGKU_b3_059_4.jpeg -->
 +
<!-- IGKU_b3_059_5.jpeg -->
 +
<!-- IGKU_b3_059_6.jpeg -->
 +
<!-- IGKU_b3_059_7.jpeg -->
 +
<!-- IGKU_b3_059_8.jpeg -->
 +
<!-- IGKU_b3_059_9.jpeg -->
 +
</div>
 +
<!------------ Table END ------------>
 +
 
 
<!-- ############################################################## Book3 Tables END  ################################################## -->
 
<!-- ############################################################## Book3 Tables END  ################################################## -->
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Latest revision as of 01:02, 2 November 2017

Notebook

July
1 2 3 4 5
6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 29 30 31
August
1 2 3 4 5
6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 29 30 31
September
1 2 3 4 5
6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 29 30 31
October
1 2 3 4 5
6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 29 30 31

07/10

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1shortAK2shortAK2B.xylophilus cDNA
2Top1Top1B.xylophilus cDNA
3long AK2long AK2B.xylophilus cDNA
4eef-1geef-1gB.xylophilus cDNA
5AK1AK1B.xylophilus cDNA
6AsbAsbB.xylophilus cDNA
714-3-3zeta14-3-3zetaB.xylophilus cDNA
8tropomiosintropomiosinB.xylophilus cDNA
914-3-3 protein14-3-3 proteinB.xylophilus cDNA

PCR Polymerase : 5*Q5 Polymerase
98.0°C : 30sec
98.0°C : 10sec
58.0°C : 30sec
72.0°C : 1min
72.0°C : 1min
4.0°C : Infinate
X35 cycle
PCR pulification: lane 2~9
Gel extraction: lane 2~9

07/11

Miniprep_Nanodrop

 Yoshimoto
sampleSample name260/280260/230ng/ul
12no datano data237.8
23no datano data180.5
34no datano data185.6
45no datano data167.2
56no datano data328.2
67no datano data133.7
78no datano data168.4
89no datano data242.5

The way of extraction : Gel extraction

07/11

Ligation

 Yoshimoto
sampleVector nameVector amount(ul)Insert nameInsert amount(ul)
1YEplac195/sma11Template21.0
2YEplac195/sma11Template31.0
3YEplac195/sma11Template41.0
4YEplac195/sma11Template51.0
5YEplac195/sma11Template61.0
6YEplac195/sma11Template71.0
7YEplac195/sma11Template81.0
8YEplac195/sma11Template91.0

07/11

Transformation

 Yoshimoto
sampleCompetentCell nameDNA name
1DH5a7/11 ligation product

07/12

Ligation

 Yoshimoto
sampleVector nameVector amount(ul)Insert nameInsert amount(ul)
1pRS3151Template1.0

07/12

Transformation

 Yoshimoto
sampleCompetentCell nameDNA name
1DH5a7/12 ligation product 2
2DH5a7/12 ligation product 3
3DH5a7/12 ligation product 4
4DH5a7/12 ligation product 5
5DH5a7/12 ligation product 6
6DH5a7/12 ligation product 7
7DH5a7/12 ligation product 8
8DH5a7/12 ligation product 9

07/13

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1T7Top1T7Top17/10 gel extraction product2
2T7long AK2T7long AK27/10 gel extraction product3
3T7eef-1gT7eef-1g7/10 gel extraction product4
4T7AK1T7AK17/10 gel extraction product5
5T7AsbT7Asb7/10 gel extraction product6
6T714-3-3zetaT714-3-3zeta7/10 gel extraction product7
7T7tropomiosinT7tropomiosin7/10 gel extraction product8
8T714-3-3 proteinT714-3-3 protein7/10 gel extraction product9

PCR Polymerase : ExTaq
98.0°C : 1min
98.0°C : 10sec
58.0°C : 30sec
72.0°C : 1min
72.0°C : 1min
4.0°C : Infinate
X35 cycle

07/14

Miniprep_Nanodrop

 Yoshimoto
sampleSample name260/280260/230ng/ul
111.841.9698.4
221.791.8576.2
331.852.1102.1
441.620.499.5
551.911.9183.6
661.821.7348.0
771.851.8583.0
881.862.8481.5
991.570.3322.6
10101.540.5517.8
11111.832.0288.0
1212.01.852.9482.0
1313.01.831.8390.6
1414.01.871.6546.2
1515.01.892.075.8
1616.01.671.98114.0
1717.01.881.4688.5
1818.01.851.9199.4
1919.01.791.7953.8
2020.01.370.5817.0
2121.01.630.570.57
2222.01.831.8453.0

The way of extraction : promega

07/15

Restriction Enzyme digestion

 Yoshimoto
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)RE2 nameRE2 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
115.0Pst10.05Sac10.0510xM1.010.0
225.0Pst10.05Sac10.0510xM1.010.0
335.0Pst10.05Sac10.0510xM1.010.0
445.0Pst10.05Sac10.0510xM1.010.0
555.0Pst10.05Sac10.0510xM1.010.0
665.0Pst10.05Sac10.0510xM1.010.0
775.0Pst10.05Sac10.0510xM1.010.0
885.0Pst10.05Sac10.0510xM1.010.0
995.0Pst10.05Sac10.0510xM1.010.0
10105.0Pst10.05Sac10.0510xM1.010.0
11115.0Pst10.05Sac10.0510xM1.010.0
12125.0Pst10.05Sac10.0510xM1.010.0
13135.0Pst10.05Sac10.0510xM1.010.0
14145.0Pst10.05Sac10.0510xM1.010.0
15155.0Pst10.05Sac10.0510xM1.010.0
16165.0Pst10.05Sac10.0510xM1.010.0
17175.0Pst10.05Sac10.0510xM1.010.0
18185.0Pst10.05Sac10.0510xM1.010.0
19195.0Pst10.05Sac10.0510xM1.010.0
20205.0Pst10.05Sac10.0510xM1.010.0
21215.0Pst10.05Sac10.0510xM1.010.0
22225.0Pst10.05Sac10.0510xM1.010.0

07/16

Miniprep_Nanodrop

 Yoshimoto
sampleSample name260/280260/230ng/ul
1T7 31.862.28255.7
2T7 41.832.13121.9
3T7 51.832.29224.2
4T7 61.852.18146.1
5T7 71.852.2596.5
6T7 81.852.27165.4
7T7 9No dataNo dataNo data

The way of extraction : promega

07/16

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1short AK2short AK2B.xylophilus cDNA
2Top1Top1B.xylophilus cDNA

PCR Polymerase : Extaq
94.0°C : 1min
94.0°C : 30sec
60.0°C : 1min
72.0°C : 2min
72.0°C : 5min
4.0°C : Infinate
X35 cycle

08/06

Nebuilder

 Yoshimoto
SampleVectornameVector amount (ul) Insert1 name Insert1 amount(ul)Insert2 name Insert2 amount (ul)Mastermix amount (ul)Total amount(ul)
1YEplac195/sma12.5Gal-AK0.8Ter-AK1.010.020.0
2YEplac195/sma12.5GPD-AK1.2Ter-AK1.010.020.0

08/06

Transformation

 Yoshimoto
sampleCompetentCell nameDNA name
1DH5aInsert

08/07

Miniprep_Nanodrop

 Yoshimoto
sampleSample name260/280260/230ng/ul
111.972.0336.0
222.031.8928.7
331.921.9834.3
441.961.8835.5
552.031.8716.2
662.012.1150.0
771.951.9530.3
881.952.0638.2
992.121.9417.0
1010no datano datano data
11111.991.9227.9
12121.961.7919.1
13131.951.9629.4
14141.981.6422.7

The way of extraction : fastgene plasmid mini kit

08/08

Transformation

 Yoshimoto
sampleCompetentCell nameDNA name
1DH5a8/6 Nebuilder product
2DH5a8/6 Nebuilder product
3DH5a8/6 Nebuilder product
4DH5a8/6 Nebuilder product
5DH5a8/6 Nebuilder product
6DH5a8/6 Nebuilder product
7DH5a8/6 Nebuilder product
8DH5a8/6 Nebuilder product

08/08

Restriction Enzyme digestion

 Yoshimoto
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
1miniprep 15.0Pst10.0510xH1.010.0
2miniprep 25.0Pst10.0510xH1.010.0
3miniprep 35.0Pst10.0510xH1.010.0
4miniprep 45.0Pst10.0510xH1.010.0
5miniprep 55.0Pst10.0510xH1.010.0
6miniprep 75.0Pst10.0510xH1.010.0

08/09

Miniprep_Nanodrop

 Yoshimoto
sampleSample name260/280260/230ng/ul
181.691.3212.1
291.971.4517.0
3111.830.9912.5
4121.921.0512.3

The way of extraction : promega PureYield Plasmid Midiprepkit

08/09

Restriction Enzyme digestion

 Yoshimoto
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
18/8 miniprep85.0Pst10.0510xH1.010.0
28/8 miniprep95.0Pst10.0510xH1.010.0
38/8 miniprep115.0Pst10.0510xH1.010.0
48/8 miniprep125.0Pst10.0510xH1.010.0

08/09

Restriction Enzyme digestion

 Fukuda
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)RE2 nameRE2 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
1AK25.0Xba10.5EcoR10.510xM6.036.0
2AK25.0Xba10.5EcoR10.510xM6.036.0
3eef-1g5.0Xba10.5EcoR10.510xM6.036.0
4eef-1g5.0Xba10.5EcoR10.510xM6.036.0
5AK15.0Xba10.5EcoR10.510xM6.036.0
6AK15.0Xba10.5EcoR10.510xM6.036.0
7Asb5.0Xba10.5EcoR10.510xM6.036.0
8Asb5.0Xba10.5EcoR10.510xM6.036.0
914-3-3-zeta5.0Xba10.5EcoR10.510xM6.036.0
1014-3-3-zeta5.0Xba10.5EcoR10.510xM6.036.0
11tropomiosin5.0Xba10.5EcoR10.510xM6.036.0
12tropomiosin5.0Xba10.5EcoR10.510xM6.036.0
1314-3-3 protein5.0Xba10.5EcoR10.510xM6.036.0
1414-3-3 protein5.0Xba10.5EcoR10.510xM6.036.0

08/10

Nebuilder

 Yoshimoto
SampleVectornameVector amount (ul) Insert1 name Insert1 amount(ul)Insert2 name Insert2 amount (ul)Mastermix amount (ul)Total amount(ul)
1YEplac195/sma12.5Gal-AK0.8Ter-AK1.010.020.0
2YEplac195/sma12.5GPD-AK1.2Ter-AK1.010.020.0

08/10

Transformation

 Yoshimoto
sampleCompetentCell nameDNA name
1DH5ainsert

08/10

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1T7AK2FT7AK2RB.xylophils cDNA1
2T7AK2FT7AK2RB.xylophils cDNA2
3T7Egf-1gFT7Egf-1gRB.xylophils cDNA3
4T7Egf-1gFT7Egf-1gRB.xylophils cDNA4
5T7AK1FT7Ak1RB.xylophils cDNA5
6T7AK1FT7Ak1RB.xylophils cDNA6
7T714-3-3zetaFT714-3-3 zetaRB.xylophils cDNA7
8T714-3-3zetaFT714-3-3 zetaRB.xylophils cDNA8
9T7tropomiosin FT7tropomiosin RB.xylophils cDNA9
10T7tropomiosin FT7tropomiosin RB.xylophils cDNA10
11T714-3-3 FT714-3-3 RB.xylophils cDNA11
12T714-3-3 FT714-3-3 RB.xylophils cDNA12
13T7AK2FT7AK2Rwater

PCR Polymerase : gotaq
94.0°C : 2min
94.0°C : 30sec
62.0°C : 30sec
68.0°C : 1min20sec
68.0°C : 7min
4.0°C : Infinate
X30 cycle

08/11

colony_PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1M13FM13R1
2M13FM13R3
3M13FM13R5
4M13FM13R7
5M13FM13R9
6M13FM13R11
7M13FM13R13
8M13FM13R15
9M13FM13R17
10M13FM13R19
11M13FM13R21
12M13FM13R23
13M13FM13R25
14M13FM13R27
15M13FM13R29
16M13FM13R31
17M13FM13R33
18M13FM13R35
19M13FM13R37
20M13FM13R39
21M13FM13R41
22M13FM13R43
23M13FM13R45
24M13FM13R47
25M13FM13R49

PCR Polymerase : QuickTaq
94.0°C : 2min
94.0°C : 30sec
60.0°C : 30sec
72.0°C : 1min
72.0°C : 1min
4.0°C : Infinate
X35 cycle

08/11

Restriction Enzyme digestion

 Ohara
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
11-15.0Pst11.010xH1.03.0
21-25.0Pst11.010xH1.03.0
31-35.0Pst11.010xH1.03.0
41-45.0Pst11.010xH1.03.0
51-55.0Pst11.010xH1.03.0
62-15.0Pst11.010xH1.03.0
71-65.0Pst11.010xH1.03.0
82-25.0Pst11.010xH1.03.0
92-35.0Pst11.010xH1.03.0
102-45.0Pst11.010xH1.03.0
112-55.0Pst11.010xH1.03.0
122-65.0Pst11.010xH1.03.0
133-15.0Pst11.010xH1.03.0
143-25.0Pst11.010xH1.03.0
153-35.0Pst11.010xH1.03.0
163-45.0Pst11.010xH1.03.0

08/11

Miniprep_Nanodrop

 Yoshimoto
sampleSample name260/280260/230ng/ul
111.881.6116.8
281.91.67170.0
391.911.71167.6
4121.881.6135.6

The way of extraction : promega PureYield Plasmid Midiprepkit

08/11

colony_PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1M13FM13R2
2M13FM13R4
3M13FM13R6
4M13FM13R8
5M13FM13R10
6M13FM13R12
7M13FM13R14
8M13FM13R16
9M13FM13R18
10M13FM13R20
11M13FM13R22
12M13FM13R24
13M13FM13R26
14M13FM13R28
15M13FM13R30
16M13FM13R32
17M13FM13R34
18M13FM13R36
19M13FM13R38
20M13FM13R40
21M13FM13R42
22M13FM13R44
23M13FM13R46
24M13FM13R48
25M13FM13R50
26M13FM13R53
27M13FM13R54
28M13FM13R55
29M13FM13R56

PCR Polymerase : Quick_taq
94.0°C : 2min
94.0°C : 30sec
60.0°C : 30sec
72.0°C : 1min
72.0°C : 10min
4.0°C : Infinate
X35 cycle

08/11

Ligation

 Yoshimoto
sampleVector nameVector amount(ul)Insert nameInsert amount(ul)
1YEplac195/sma11AK2-gal11.0
2YEplac195/sma11AK2-GPD1.0

08/11

Restriction Enzyme digestion

 Ohara
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
17A5.0Pst10.510xH6.060.0
27B5.0Pst10.510xH6.060.0

08/11

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1M13FM13RAK2-gal1 Nebuilder
2M13FM13RAK2 GPD Nebuilder
3M13FM13Rgfp gal Nebuilder
4M13FM13Rgfp GPD Nebuilder
5M13FM13RAK1 gal Nebuilder
6M13FM13RAK1 GPD Nebuilder

PCR Polymerase : 2*KAPA 2G Robust HS Readymix
94.0°C : 2min
94.0°C : 15sec
60.0°C : 15sec
72.0°C : 40sec
72.0°C : 1min
4.0°C : Infinate
X35 cycle

08/11

Transformation

 Yoshimoto
sampleCompetentCell nameDNA name
1DH5ainsert

08/12

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1AK1FAK1R8
2AK1FAK1R9
3AK1FAK1R12
4AK1FAK1R8/6 gal
5AK1FAK1R8/6 GPD
6AK1FAK1Rgal1 AK1
7AK1FAK1Rgal1 AK1
8AK1FAK1Rwater

PCR Polymerase : quicktaq
94.0°C : 2min
94.0°C : 30sec
60.0°C : 30sec
68.0°C : 1min
68.0°C : 7min
4.0°C : Infinate
X35 cycle

08/12

Ligation

 Yoshimoto
sampleVector nameVector amount(ul)Insert nameInsert amount(ul)
1YEplac195/sma11gblock 711.0
2YEplac195/sma11gblock 721.0
3YEplac195/sma11gblock 731.0
4YEplac195/sma11gblock 741.0
5YEplac195/sma11gblock 751.0
6YEplac195/sma11gblock 761.0
7YEplac195/sma11gblock 771.0
8YEplac195/sma11gblock 781.0

08/12

Transformation

 Yoshimito
sampleCompetentCell nameDNA name
1DH5ainsert

08/13

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1AK2FAK2F8
2AK2FAK2F9
3AK2FAK2F12
4AK2FAK2F8/6 gal
5AK2FAK2F8/6 GPD
6AK2FAK2F7
7AK2FAK2Fwater

PCR Polymerase : 2*KAPA hifi HS ready mix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 2min
72.0°C : 2min
4.0°C : Infinate
X40 cycle

08/13

Restriction Enzyme digestion

 Ohara
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
115.0Pst11.010xH1.010.0
225.0Pst11.010xH1.010.0
335.0Pst11.010xH1.010.0
445.0Pst11.010xH1.010.0
555.0Pst11.010xH1.010.0
675.0Pst11.010xH1.010.0
715.0Pst11.010xH1.010.0
875.0Pst11.010xH1.010.0
995.0Pst11.010xH1.010.0
1085.0Pst11.010xH1.010.0
11125.0Pst11.010xH1.010.0

08/13

Restriction Enzyme digestion

 Ohara
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
1145.0Pst11.010xH1.010.0
2165.0Pst11.010xH1.010.0
3225.0Pst11.010xH1.010.0
4245.0Pst11.010xH1.010.0
5265.0Pst11.010xH1.010.0
6305.0Pst11.010xH1.010.0
7365.0Pst11.010xH1.010.0
8245.0Pst11.010xH1.010.0
9145.0Pst10.010xH0.010.0
10225.0Pst10.010xH0.010.0
11365.0Pst10.010xH0.010.0

08/13

colony PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1M13FM13R77-1
2M13FM13R77-2
3M13FM13R77-3
4M13FM13R78-4
5M13FM13R78-5

PCR Polymerase : 2*KAPA hifi HS ready mix
95.0°C : 1min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min
72.0°C : 1min
4.0°C : Infinate
X35 cycle

08/13

Miniprep_Nanodrop

 Yoshimoto
sampleSample name260/280260/230ng/ul
1141.911.4482.6
2161.961.6791.4
3222.041.7796.7
4241.881.593.0
5261.891.5189.4
6301.91.4874.6
7361.911.587.5

The way of extraction : promega PureYield Plasmid Miniprepkit

08/13

Ligation

 Yoshimoto
sampleVector nameVector amount(ul)Insert nameInsert amount(ul)
1YEplac195/sma11Gal-AK2 992.0
2YEplac195/sma11GPD-AK2 802.0
3YEplac195/sma11Ter-AK2 812.0
4YEplac195/sma11T7 AK1 822.0
5YEplac195/sma11T7 AK1.2 832.0
6YEplac195/sma11Gal1 gfp 852.0
7YEplac195/sma11ter gfp 862.0

08/13

Nebuilder

 Yoshimoto
SampleVectornameVector amount (ul) Insert1 name Insert1 amount(ul)Insert2 name Insert2 amount (ul)Mastermix amount (ul)Total amount(ul)
1YEPlac195/sma10.25GPD-AK1.5Ter-AK5.05.012.0
2YEPlac195/sma10.25Gal-AK2.0Ter-AK5.05.012.0

08/13

Transformation

 Yoshimoto
sampleCompetentCell nameDNA name
1DH5aNebuilder product
2DH5aNebuilder product
3DH5aNebuilder product
4DH5aNebuilder product

08/14

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1M13FM13R79-1
2M13FM13R80-1
3M13FM13R80-2
4M13FM13R81-1
5M13FM13R97-1
6M13FM13R98-1
7M13FM13R98-2
8M13FM13R98-3
9M13FM13R92-1
10M13FM13R99-1
11M13FM13R100-1
12M13FM13R101-1
13M13FM13R101-2
14M13FM13R101-3
15M13FM13R102-1
16M13FM13Rwater

PCR Polymerase : 2*KAPA hifi HS readymix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min20sec
72.0°C : 1min
4.0°C : Infinate
X35 cycle

08/14

Restriction Enzyme digestion

 Ohara
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)RE2 nameRE2 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
125.0Pst11.0NoneNone10xH1.010.0
235.0Pst11.0NoneNone10xH1.010.0
345.0Pst11.0NoneNone10xH1.010.0
455.0Pst11.0NoneNone10xH1.010.0
575.0Pst11.0NoneNone10xH1.010.0
6145.0Pst11.0NoneNone10xH1.010.0
7165.0Pst11.0NoneNone10xH1.010.0
8225.0Pst11.0NoneNone10xH1.010.0
977-15.0Pst11.0EcoR11.010xH1.010.0
1077-25.0Pst11.0EcoR11.010xH1.010.0
1177-35.0Pst11.0EcoR11.010xH1.010.0
12245.0Pst11.0NoneNone10xH1.010.0
13265.0Pst11.0NoneNone10xH1.010.0
14305.0Pst11.0NoneNone10xH1.010.0
15365.0Pst11.0NoneNone10xH1.010.0
1678-15.0Pst11.0NoneNone10xH1.010.0
1778-25.0Pst11.0NoneNone10xH1.010.0
18785.0Pst11.0NoneNone10xH1.010.0
19995.0Pst11.0NoneNone10xH1.010.0
201005.0Pst11.0NoneNone10xH1.010.0
211015.0Pst11.0NoneNone10xH1.010.0

08/14

colony_PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1M13FM13R97
2M13FM13R98
3M13FM13R99
4M13FM13R100
5M13FM13R101
6M13FM13R102

PCR Polymerase : 2*KAPA hifi HS readymix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min20sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

08/14

Transformation

 Yoshimoto
sampleCompetentCell nameDNA name
1DH5aNebuilder product gal1-gfp
2DH5aNebuilder product GPD-gfp
3DH5a8/13 ligation product Gal-AK2
4DH5a8/13 ligation product GPD-AK2
5DH5a8/13 ligation product Ter-AK2

08/14

Transformation

 Yoshimoto
sampleCompetentCell nameDNA name
1DH5aligation product Gal-AK2
2DH5aligation product GPD-AK2
3DH5aligation product Ter-AK2

08/14

Miniprep_Nanodrop

 Yoshimoto
sampleSample name260/280260/230ng/ul
11011.970.9324.9
21001.990.9934.8
3971.941.3550.1
4981.971.9740.3
599.01.961.6599.7
673.51.820.947.9
778.41.860.9943.7
871-31.881.2167.4
9361.881.41129.5
1077-11.810.9970.8
1177-21.891.263.0
12301.821.23135.8
13261.861.31143.9
14241.881.43141.5
15141.821.31178.7
16161.911.37132.3
17221.791.35207.8
1871.851.2894.4
1951.790.9163.8
2041.660.8164.2
2121.870.9650.5
2231.791.1567.1

The way of extraction : promega

08/15

Restriction Enzyme digestion

 Ohara
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
1925.0Pst11.010xH1.010.0
2975.0Pst11.010xH1.010.0
3985.0Pst11.010xH1.010.0
41005.0Pst11.010xH1.010.0
51015.0Pst11.010xH1.010.0
6925.0Pst11.010xH1.010.0
7975.0Pst11.010xH1.010.0
8985.0Pst11.010xH1.010.0
91005.0Pst11.010xH1.010.0
101015.0Pst11.010xH1.010.0

08/15

Nebuilder

 Yoshimoto
SampleVectornameVector amount (ul) Insert1 name Insert1 amount(ul)Insert2 name Insert2 amount (ul)Mastermix amount (ul)Total amount(ul)
1YEplac195/sma10.25Gal-AK1.825Ter-AK1.55.010.0
2YEplac195/sma10.25GPD-AK1.25Ter-AK1.55.010.0

08/15

Transformation

 Yoshimoto
sampleCompetentCell nameDNA name
1DH5a111
2DH5a112

08/17

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK25IK26Rev
2IK27IK22yeast genome
3IK23IK28yeast genome

PCR Polymerase : 2*KAPA hifi HS Ready mix
98.0°C : 3min
98.0°C : 20sec
62.0°C : 30sec
72.0°C : 1min
72.0°C : 5min
4.0°C : Infinate
X35 cycle

08/17

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK27IK22yeast genome
2IK23IK28yeast genome
3IK27IK28GPDp+Rev+GPDt

PCR Polymerase : 2*KAPA hifi HS ready mix
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 1min
72.0°C : 5min
4.0°C : Infinate
X35 cycle

08/17

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK25IK26Rev
2IK27IK22yeast genome
3IK23IK28yeast genome
4IK25IK26Rev after purification
5IK27IK22GPDp after purification
6IK23IK28GPDt after purification

PCR Polymerase : 2*KAPA hifi HS ready mix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X36 cycle

08/17

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1M13FM13R86-1
2M13FM13R86-2
3M13FM13R81-1
4M13FM13R81-2
5M13FM13R81-3
6M13FM13R95-1
7M13FM13R95-2
8M13FM13R95-3
9M13FM13R95-4
10M13FM13R93-1
11M13FM13R93-2
12M13FM13R92
13M13FM13Rbrue colony
14M13FM13R11

PCR Polymerase : 2*KAPA hifi HS readymix
95.0°C : 3min
95.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min20sec
72.0°C : 1min
4.0°C : Infinate
X35 cycle

08/17

Nebuilder

 Yoshimoto
SampleVectornameVector amount (ul) Insert1 name Insert1 amount(ul)Insert2 name Insert2 amount (ul)Mastermix amount (ul)Total amount(ul)
1YEplac195/sma10.8Gal-AK26.5Ter-AK25.010.022.3
2YEplac195/sma10.8GPD-AK24.2Ter-AK25.010.020.0

08/17

Transformation

 Yoshimoto
sampleCompetentCell nameDNA name
1BL21113
2BL21114
3BL21115

08/18

Restriction Enzyme digestion

 Ohara
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)RE2 nameRE2 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
179-25.0Pst11.0EcoR11.010xH1.010.0
279-35.0Pst11.0EcoR11.010xH1.010.0
379-65.0Pst11.0EcoR11.010xH1.010.0
480-25.0Pst11.0EcoR11.010xH1.010.0
580-75.0Pst11.0EcoR11.010xH1.010.0
680-135.0Pst11.0EcoR11.010xH1.010.0
781-15.0Pst11.0NoneNone10xH1.010.0
881-25.0Pst11.0NoneNone10xH1.010.0
9975.0Pst11.0NoneNone10xH1.010.0
1099-A5.0Pst11.0NoneNone10xH1.010.0
1199-B5.0Pst11.0NoneNone10xH1.010.0
121005.0Pst11.0NoneNone10xH1.010.0
13100-15.0Pst11.0NoneNone10xH1.010.0
14100-25.0Pst11.0NoneNone10xH1.010.0
15101-15.0Pst11.0NoneNone10xH1.010.0
16101 35.0Pst11.0NoneNone10xH1.010.0
171025.0Pst11.0NoneNone10xH1.010.0

08/18

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK27IK28Rev

PCR Polymerase : 2*KAPA hifi HS Ready mix
98.0°C : 3min
98.0°C : 20sec
60.0°C : 20sec
72.0°C : 1min20sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

08/18

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1M13FAK2R79-2
2M13FAK2R79-3
3M13FAK2R80-13
4M13FAK2F79-2
5M13FAK2F79-3
6M13FAK2F80-13
7M13RM13Fblue colony

PCR Polymerase : 2*KAPA hifi HS readymix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min30sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

08/18

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK22IK27GPDp after purification
2IK22IK27GPDp after purification
3IK22IK27GPDp after purification
4IK22IK27GPDp after purification
5M13FAK2R113gal
6M13FAK2R114GPD

PCR Polymerase : 2*KAPA hifi HS readymix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min30sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
PCR pulification: lane1~4
Gel extraction: lane1~4

08/19

Ligation

 Yoshimoto
sampleVector nameVector amount(ul)Insert nameInsert amount(ul)
11410.342T7akRRE3'9.4
21420.342T7ak1stem8.3
31430.341T7ak1stem4.4
41440.342T7RREak15.3
51450.341T7RREak18.5
61460.34GPD+AK213.2
71470.34Gal1+AK215.8
81480.34AK2+ter20.1

08/20

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK22IK27GPDp
2IK22IK27GPDp
3IK22IK27yeast genome
4IK22IK27yeast genome
5IK22IK27yeast genome
6IK22IK27yeast genome
7IK27IK28GPDp+Rev+GPDt
8IK27IK28GPDp+Rev+GPDt

PCR Polymerase : 2*KAPA hifi HS ready mix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min30sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

08/20

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK27IK22GPDp
2IK21IK22GPDp

PCR Polymerase : 2*KAPA HS readymix
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 1min
72.0°C : 5min
4.0°C : Infinate
X35 cycle

08/21

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK29IK30Rev
2IK29IK30Rev
3IK29IK30Rev
4IK29IK30Rev
5M13RAK1R141 1-1
6M13RAK1R141 1-2
7M13RAK1R141 1-3
8M13RAK1R141 1-4
9M13RAK1R142 1-1
10M13RAK1R142 1-2
11M13RAK1R142 1-3
12M13RAK1R142 1-4
13M13RAK1F141 1-1
14M13RAK1F141 1-2
15M13RAK1F141 1-3
16M13RAK1F141 1-4
17M13RAK1F142 1-1
18M13RAK1F142 1-2
19M13RAK1F142 1-3
20M13RAK1F142 1-4

PCR Polymerase : 2*KAPA hifi HS Readymix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min20sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

08/21

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1M13FAK1R141 2-1
2M13FAK1R141 2-4
3M13FAK1R141 1-15
4M13FAK1R142 1-5
5M13FAK1R143 9
6M13FAK1R143 13
7M13FAK1R143 27
8M13FAK1R143 2
9M13FAK1F141 2-1
10M13FAK1F141 2-4
11M13FAK1F141 1-15
12M13FAK1F142 1-5
13M13FAK1F143 9
14M13FAK1F143 13
15M13FAK1F143 27
16M13FAK1F143 2

PCR Polymerase : 2*KAPA hifi HS Readymix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min20sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

08/21

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK29IK30Rev
2IK29IK30Rev
3IK29IK30Rev
4IK29IK30Rev

PCR Polymerase : 2*KAPA hifi HS Readymix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min20sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
PCR pulification: lane2~4
Gel extraction: lane2~4

08/22

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK21IK22GPDp
2IK21IK22GPDp
3IK21IK22GPDp
4IK21IK22GPDp
5IK21IK22water
6IK23IK28GPDt
7IK23IK28GPDt
8IK23IK28GPDt
9IK23IK28GPDt
10IK23IK28water

PCR Polymerase : 2*KAPA hifi HS Readymix
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 1min
72.0°C : 5min
4.0°C : Infinate
X35 cycle
PCR pulification: lane2~4
Gel extraction: lane2~4

08/22

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK27IK28GPDp+Rev+GPDt
2IK27IK28GPDp+Rev+GPDt
3IK23IK24yeast genome
4IK23IK24yeast genome
5IK23IK24yeast genome
6IK23IK24yeast genome
7IK21IK24GPDp+Rev+GPDt

PCR Polymerase : 2*KAPA hifi HS ReadyMIx
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min30sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

08/22

PCR

 Fukuda
lanePrimer FowardPrimer ReveseTemplate
1M13FM13R144 11
2M13FM13R144 3
3M13FM13R144 7
4M13FM13R144 1
5M13FM13R144 17
6M13FM13R144 5
7M13FM13R144 9
8M13FM13R144 13
9M13FM13R142 1
10M13FM13R142 5
11M13FM13R142 3
12M13FM13R142 4
13M13FM13R142 2-1
14M13FM13R142 2-2
15M13FM13R142 2-3
16M13FM13R142 2-4
17M13FM13R143 13
18M13FM13R143 15
19M13FM13R143 5
20M13FM13R153 21
21M13FM13R143 17
22M13FM13R143 19
23M13FM13R143 25
24M13FM13R143 9

PCR Polymerase : 2*KAPA hifi HS ReadyMix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min30sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

08/22

PCR

 Takimoto
lanePrimer FowardPrimer ReveseTemplate
1M13FM13R145 1-13
2M13FM13R145 2-13
3M13FM13R145 2-15
4M13FM13R145 1-5
5M13FM13R145 2-17
6M13FM13R145 2-11
7M13FM13R145 2-9
8M13FM13R145 1-17
9M13FM13R146 2-3
10M13FM13R146 2-1
11M13FM13R146 1-3
12M13FM13R146 2-7
13M13FM13R146 2-9
14M13FM13R146 2-5
15M13FM13R146 1-4

PCR Polymerase : 2*KAPA hifi HS ReadyMix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min30sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

08/22

Nebuilder

 Yoshimoto
SampleVectornameVector amount (ul) Insert1 name Insert1 amount(ul)Insert2 name Insert2 amount (ul)Mastermix amount (ul)Total amount(ul)
1171.0Vector amount (ul)Gal1+gfp-20.78Gal1+gfp1.0310.0Total amount(ul)
2172.0Vector amount (ul)Gal1+AK21.26Gal1+AK2-21.0110.0Total amount(ul)
3173.0Vector amount (ul)164.00.65165.00.5710.0Total amount(ul)
4174.0Vector amount (ul)161.00.65162.00.7610.0Total amount(ul)
5175.0Vector amount (ul)161.00.38167.00.8310.0Total amount(ul)
6176.0Vector amount (ul)166.00.38165.00.8310.0Total amount(ul)
7177.0Vector amount (ul)163.00.38162.00.8310.0Total amount(ul)
8178.0Vector amount (ul)163.00.38167.00.8310.0Total amount(ul)

08/22

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1NoneNoneGPDp+Rev+GPDt
2IK27IK28lane1 after 15 cycle

PCR Polymerase : 2*KAPA hifi Hs ReadyMix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min30sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

08/22

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK23IK28GPDt
2IK23IK28GPDt
3IK23IK28GPDt
4IK23IK28GPDt
5IK27IK30GPDp + Rev
6IK21IK30GPDp + Rev

PCR Polymerase : 2*KAPA hifi Robust
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

08/23

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1M13FM13R147 2-17
2M13FM13R147 1-2
3M13FM13R147 2-3
4M13FM13R147 7
5M13FM13R141 1-1
6M13FM13R141 1-2
7M13FM13R141 1-4
8M13FM13R148 2-3
9M13FM13R148 2-5
10M13FM13R148 2-4
11M13FM13R148 1-5
12M13FM13R148 1-3
13M13FM13R144 11
14M13FM13R144 3
15M13FM13R144 1
16M13FM13R146 1-6
17M13FM13R146 1-4
18M13FM13R146 2-5
19M13FM13R146 2-7
20M13FM13R142 2-2
21M13FM13R142 1-1
22M13FM13R142 2-7
23M13FM13R142 2-3
24M13FM13R145 2-11
25M13FM13R145 1-17
26M13FM13R145 2-9
27M13FM13R145 2-17
28M13FM13R143 5
29M13FM13R143 17
30M13FM13R143 15
31M13FM13R143 21
32M13FAK1R147 2-17
33M13FAK1R147 1-2
34M13FAK1R147 2-3
35M13FAK1R147 7
36M13FAK1R141 1-1
37M13FAK1R141 1-2
38M13FAK1R141 1-4
39M13FAK1R148 2-3
40M13FAK1R148 2-5
41M13FAK1R148 2-4
42M13FAK1R148 1-5
43M13FAK1R148 1-3
44M13FAK1R144 11
45M13FAK1R144 3
46M13FAK1R144 1
47M13FAK1R146 1-6
48M13FAK1R146 1-4
49M13FAK1R146 2-5
50M13FAK1R146 2-7
51M13FAK1R142 2-2
52M13FAK1R142 1-1
53M13FAK1R142 2-7
54M13FAK1R142 2-3
55M13FAK1R145 2-11
56M13FAK1R145 1-17
57M13FAK1R145 2-9
58M13FAK1R145 2-17
59M13FAK1R143 5
60M13FAK1R143 17
61M13FAK1R143 15
62M13FAK1R143 21

PCR Polymerase : 2*KAPA hifi robust
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

08/24

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1M13FM13R141 2-1
2M13FM13R141 2-2
3M13FM13R141 2-4
4M13FM13R141 2-6
5M13FM13R142 1-5
6M13FM13R142 2-4
7M13FM13R142 1-3
8M13FM13R142 1-4
9M13FM13R143 13
10M13FM13R143 19
11M13FM13R143 25
12M13FM13R144 18
13M13FM13R144 17
14M13FM13R144 5
15M13FM13R144 9
16M13FM13R143 19
17M13FM13R145 1-5
18M13FM13R145 2-15
19M13FM13R143 2-13
20M13FM13R145 1-13
21M13FAK1R141 2-1
22M13FAK1R141 2-2
23M13FAK1R141 2-4
24M13FAK1R141 2-6
25M13FAK1R142 1-5
26M13FAK1R142 2-4
27M13FAK1R142 1-3
28M13FAK1R142 1-4
29M13FAK1R145 1-5
30M13FAK1R145 2-15
31M13FAK1R143 2-13
32M13FAK1R145 1-13
33M13FAK1R144 17
34M13FAK1R144 5
35M13FAK1R144 9
36M13FAK1R143 19
37M13FAK1R143 13
38M13FAK1R143 19
39M13FAK1R143 25
40M13FAK1R144 9

PCR Polymerase : 2*KAPA Robust
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

08/26

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK27IK30GPDp + Rev after gel extraction
2IK27IK30GPDp + Rev after gel extraction
3IK27IK30GPDp + Rev after gel extraction
4IK27IK30GPDp + Rev after gel extraction
5IK23IK28GPDt after PCR purification
6IK23IK28GPDt after PCR purification
7IK23IK28GPDt after PCR purification
8IK23IK28GPDt after PCR purification

PCR Polymerase : 2*KAPA hifi HS Readymix
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

08/26

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1M13FM13R145-2 11
2M13FM13R145-1 9
3M13FM13R145 2-7
4M13FM13R145 1-21
5M13FM13R145 1-3
6M13FM13R145 2-3
7M13FM13R145 1-19
8M13FM13R142 1-2
9M13FAK1F145-2 11
10M13FAK1F145-1 9
11M13FAK1F145 2-7
12M13FAK1F145 1-21
13M13FAK1F145 1-3
14M13FAK1F145 2-3
15M13FAK1F145 1-19
16M13FAK1F142 1-2

PCR Polymerase : 2*KAPA hifi
98.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 25sec
72.0°C : 3min
4.0°C : Infinate
X35 cycle

08/26

Restriction Enzyme digestion

 Ohara
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
1114-12.0Pst11.010xH1.010.0
2114-22.0Pst11.010xH1.010.0
31722.0Pst11.010xH1.010.0
4172-12.0Pst11.010xH1.010.0
5172-22.0Pst11.010xH1.010.0
6172-42.0Pst11.010xH1.010.0
7175-32.0Pst11.010xH1.010.0
8175-102.0Pst11.010xH1.010.0
91762.0Pst11.010xH1.010.0
101772.0Pst11.010xH1.010.0

08/27

Nebuilder

 Yoshimoto
SampleVectornameVector amount (ul) Insert1 name Insert1 amount(ul)Insert2 name Insert2 amount (ul)Mastermix amount (ul)Total amount(ul)
1Rev3.02GPDp+Rev0.669GPDt0.87510.0Total amount(ul)
2173.03.02164.00.325165.00.2855.0Total amount(ul)
3175.03.02161.00.19167.00.425.0Total amount(ul)
4178.03.02163.00.41167.00.425.0Total amount(ul)

08/27

Restriction Enzyme digestion

 Ohara
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
1171-15.0Pst11.010xH1.010.0
2171-1-25.0Pst11.010xH1.010.0
3172-25.0Pst11.010xH1.010.0
4172-2-25.0Pst11.010xH1.010.0
5172-35.0Pst11.010xH1.010.0
6172-45.0Pst11.010xH1.010.0
7172-55.0Pst11.010xH1.010.0
8172-65.0Pst11.010xH1.010.0
9172-85.0Pst11.010xH1.010.0
10172-95.0Pst11.010xH1.010.0
11172-105.0Pst11.010xH1.010.0
12172-115.0Pst11.010xH1.010.0
13172-125.0Pst11.010xH1.010.0
14172-135.0Pst11.010xH1.010.0
15175-15.0EcoR11.010xH1.010.0
16175-25.0EcoR11.010xH1.010.0
17175-35.0EcoR11.010xH1.010.0
18175-45.0EcoR11.010xH1.010.0
19176-15.0EcoR11.010xH1.010.0
20177-25.0EcoR11.010xH1.010.0
21177-35.0EcoR11.010xH1.010.0
22177-45.0EcoR11.010xH1.010.0
23177-55.0EcoR11.010xH1.010.0
24177-15.0EcoR11.010xH1.010.0
25177-1-25.0EcoR11.010xH1.010.0
26171-15.0EcoR11.010xH1.010.0
27172-25.0EcoR11.010xH1.010.0
28175-15.0EcoR11.010xH1.010.0
29175-25.0EcoR11.010xH1.010.0
30175-35.0EcoR11.010xH1.010.0
31175-45.0EcoR11.010xH1.010.0
32176-15.0EcoR11.010xH1.010.0
33176-25.0EcoR11.010xH1.010.0
34177-25.0EcoR11.010xH1.010.0
35177-35.0EcoR11.010xH1.010.0
36177-45.0EcoR11.010xH1.010.0
37177-55.0EcoR11.010xH1.010.0

08/27

Restriction Enzyme digestion

 Ohara
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
1171 1-15.0Pst11.010xH1.010.0
2171 1-25.0Pst11.010xH1.010.0
3171 25.0Pst11.010xH1.010.0
4171 35.0Pst11.010xH1.010.0
5172-15.0Pst11.010xH1.010.0
6172-25.0Pst11.010xH1.010.0
7172-35.0Pst11.010xH1.010.0
8172-4-15.0Pst11.010xH1.010.0
9172-4-25.0Pst11.010xH1.010.0
10172-55.0Pst11.010xH1.010.0
11172-65.0Pst11.010xH1.010.0
12172-85.0Pst11.010xH1.010.0
13172-95.0Pst11.010xH1.010.0
14172-105.0Pst11.010xH1.010.0
15172-115.0Pst11.010xH1.010.0
16172-125.0Pst11.010xH1.010.0
17172-135.0Pst11.010xH1.010.0
18171-1-15.0EcoR11.010xH1.010.0
19174-15.0EcoR11.010xH1.010.0
20175-15.0EcoR11.010xH1.010.0
21175-25.0EcoR11.010xH1.010.0
22175-35.0EcoR11.010xH1.010.0
23175-45.0EcoR11.010xH1.010.0
24176-15.0EcoR11.010xH1.010.0
25176-25.0EcoR11.010xH1.010.0
26177-25.0EcoR11.010xH1.010.0
27177-35.0EcoR11.010xH1.010.0
28177-45.0EcoR11.010xH1.010.0
29177-55.0EcoR11.010xH1.010.0

08/28

Restriction Enzyme digestion

 Yoshimoto
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)RE2 nameRE2 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
1145-25.0Xba10.5EcoR10.510xM1.010.0
2142 2-115.0Xba10.5EcoR10.510xM1.010.0
3142 2-25.0Xba10.5EcoR10.510xM1.010.0
4142 2-15.0Xba10.5EcoR10.510xM1.010.0
5142 2-35.0Xba10.5EcoR10.510xM1.010.0
6242 1-45.0Xba10.5EcoR10.510xM1.010.0
7142 2-45.0Xba10.5EcoR10.510xM1.010.0
8142 1-55.0Xba10.5EcoR10.510xM1.010.0
9142 1-35.0Xba10.5EcoR10.510xM1.010.0
10142 2-55.0Xba10.5EcoR10.510xM1.010.0
11142 1-25.0Xba10.5EcoR10.510xM1.010.0
12142 1-15.0Xba10.5EcoR10.510xM1.010.0
13145 2-35.0Xba10.5EcoR10.510xM1.010.0
14145 1-25.0Xba10.5EcoR10.510xM1.010.0
15145 2-75.0Xba10.5EcoR10.510xM1.010.0
16145 4-95.0Xba10.5EcoR10.510xM1.010.0
17145 1-115.0Xba10.5EcoR10.510xM1.010.0
18145 1-35.0Xba10.5EcoR10.510xM1.010.0
19145 2-115.0Xba10.5EcoR10.510xM1.010.0
20145 2-55.0Xba10.5EcoR10.510xM1.010.0
21145 2-85.0Xba10.5EcoR10.510xM1.010.0
22145 1-145.0Xba10.5EcoR10.510xM1.010.0
23145 2-155.0Xba10.5EcoR10.510xM1.010.0
24145 2- 4-25.0Xba10.5EcoR10.510xM1.010.0
25145 1-175.0Xba10.5EcoR10.510xM1.010.0
26145 2-175.0Xba10.5EcoR10.510xM1.010.0

08/29

Restriction Enzyme digestion

 Yoshimoto
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)RE2 nameRE2 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
1142 1-35.0EcoR10.5Not10.510xH, BSA1ul 1ul10.0
2145 1-115.0Xba10.5Not10.510xK, BSA1ul 1ul9.0
3145 1-35.0Xba10.5Not10.510xK, BSA1ul 1ul9.0
4145 2-425.0Xba10.5Not10.510xK, BSA1ul 1ul9.0

08/29

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK27IK28Rev, GPDp, GPDt
2IK27IK28Rev+GPDp, GPDt
3IK27IK28Rev plasmid
4IK27IK28Rev plasmid
5IK27IK28Rev plasmid
6IK27IK28Rev plasmid

PCR Polymerase : KAKA Robust
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min30sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

08/31

Restriction Enzyme digestion

 Yoshimoto
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)RE2 nameRE2 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
1141 1-23.5EcoR10.5Xba10.510xN1.010.0
2141 1-23.5EcoR10.5Xba10.510xN1.010.0
3141 1-33.5EcoR10.5Xba10.510xN1.010.0
4141 1-33.5EcoR10.5Xba10.510xN1.010.0
5144-93.5EcoR10.5Not10.510xH, BSA1ul, 1ul10.0
6144-93.5EcoR10.5Not10.510xH, BSA1ul, 1ul10.0
7144 173.5EcoR10.5Not10.510xH, BSA1ul, 1ul10.0
8144 173.5EcoR10.5Not10.510xH, BSA1ul, 1ul10.0
9144 33.5EcoR10.5Not10.510xH, BSA1ul, 1ul10.0
10144 33.5EcoR10.5Not10.510xH, BSA1ul, 1ul10.0
11144 33.5EcoR10.5Not10.510xH, BSA1ul, 1ul10.0
12144 13.5EcoR10.5Not10.510xH, BSA1ul, 1ul10.0
13144 13.5EcoR10.5Not10.510xH, BSA1ul, 1ul10.0
14144 13.5EcoR10.5Not10.510xH, BSA1ul, 1ul10.0
15143 213.5Xba10.5Not10.510xK, BSA1ul, 1ul10.0
16143 213.5Xba10.5Not10.510xK, BSA1ul, 1ul10.0
17143 213.5Xba10.5Not10.510xK, BSA1ul, 1ul10.0
18143 153.5Xba10.5Not10.510xK, BSA1ul, 1ul10.0
19143 153.5Xba10.5Not10.510xK, BSA1ul, 1ul10.0
20143 153.5Xba10.5Not10.510xK, BSA1ul, 1ul10.0
21143 153.5Xba10.5Not10.510xK, BSA1ul, 1ul10.0
22143 53.5Xba10.5Not10.510xK, BSA1ul, 1ul10.0
23143 53.5Xba10.5Not10.510xK, BSA1ul, 1ul10.0
24143 53.5Xba10.5Not10.510xK, BSA1ul, 1ul10.0
25143 133.5Xba10.5Not10.510xK, BSA1ul, 1ul10.0
26145 1-33.5Xba10.5Not10.510xK, BSA1ul, 1ul10.0
27145 1-33.5Xba10.5Not10.510xK, BSA1ul, 1ul10.0
28145 1-33.5Xba10.5Not10.510xK, BSA1ul, 1ul10.0
29142 2-25.0Xba10.5EcoR10.510xM1.010.0
30142 1-35.0Xba10.5EcoR10.510xM1.010.0
311425.0Xba10.5EcoR10.510xM1.010.0
32142 2-25.0Not10.5Xba10.510xK BSA1ul, 1ul10.0
33142 1-35.0Not10.5Xba10.510xK BSA1ul, 1ul10.0
341425.0Not10.5Xba10.510xK BSA1ul, 1ul10.0

09/01

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1M13FM13RP1
2M13FM13RP2
3M13FM13RP3
4M13FM13RP4
5M13FM13RT1
6M13FM13RT2
7M13FM13RT3
8M13FM13RT4

PCR Polymerase : KAPA
98.0°C : 3min
98.0°C : 30sec
60.0°C : 20sec
72.0°C : 40sec
72.0°C : 10min
4.0°C : Infinate
X35 cycle

09/02

Restriction Enzyme digestion

 Yoshimoto
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)RE2 nameRE2 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
1144-33.5Xba10.5EcoR10.510xM1.010.0
21423.5Xba10.5EcoR10.510xM1.010.0
3145 1-113.5Xba10.5EcoR10.510xM1.010.0

09/02

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1M13FM13R165-1
2M13FM13R165-2
3M13FM13R165-3
4M13FM13R165-4
5M13FIK60165-1
6M13FIK60165-2
7M13FIK60165-3
8M13FIK60165-4

PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 30sec
60.0°C : 20sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/02

Restriction Enzyme digestion

 Yoshimoto
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)RE2 nameRE2 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
1142 2-23.5Xba10.5EcoR10.510xM1.010.0
214213.5Xba10.5EcoR10.510xM1.010.0
3142 1-33.5Xba10.5EcoR10.510xM1.010.0

09/03

Ligation

 Yoshimoto
sampleVector nameVector amount(ul)Insert nameInsert amount(ul)
1YEplac195/sma10.351616.19
2YEplac195/sma10.3516212.2
3YEplac195/sma10.3516313.1
4YEplac195/sma10.3516410.32
5YEplac195/sma10.3516617.25
6YEplac195/sma10.3516713.28

09/03

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1M13FIK55GFPgalp-1
2M13FIK55GFPgalp-2
3M13FIK55GFPgalp-3
4M13FIK55GFPgalp-4
5M13FIK55GFPt-1
6M13FIK55GFPt-2
7M13FIK55GFPt-3
8M13FIK55GFPt-4

PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 30sec
60.0°C : 20sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/04

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK21IK22GPDp
2IK21IK22GPDp
3IK21IK22GPDp
4IK21IK22GPDp
5IK23IK28GPDter
6IK23IK28GPDter
7IK23IK28GPDter
8IK23IK28GPDter
9IK29IK30Rev long
10IK29IK30Rev long
11IK29IK30Rev long
12IK29IK30Rev long
13IK21IK28GPDter(PCRp), Rev(PCRp), GPDp(gel ex)
14IK21IK28GPDter(PCRp), Rev+GPDp(PCRp)

PCR Polymerase : KAPA
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 1min20sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
PCR pulification: 1~4
PCR pulification: 9~12
Gel extraction: 1~4
Gel extraction: 9~12

09/04

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK60AK1F142-2-1
2IK60AK1F142-2-2
3IK60AK1F142-2-3
4IK60AK1F142-2-4
5IK60AK1F142-2-5
6IK60AK1F142-1-1
7IK60AK1F142-1-2
8IK60AK1F142-1-3
9IK60AK1F142-1-4
10IK60AK1F142-1-5
11IK60M13F144-3
12IK60M13R145-1-11
13IK60AK1F144-3
14IK60AK1F145-1-11

PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 30sec
60.0°C : 20sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/04

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK60M13R142-2-1
2IK60M13R142-2-2
3IK60M13R142-2-3
4IK60M13R142-2-4
5IK60M13R142-2-5
6IK60M13R142-1-1
7IK60M13R142-1-2
8IK60M13R142-1-3
9IK60M13R142-1-4
10IK60M13R142-1-5
11M13FM13Rgfpp
12IK55M13Rgfpp
13M13FM13Rgfpt M13
14IK55M13Rgfp 55

PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 20sec
61.0°C : 20sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/04

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK27IK30Rev+p
2IK21IK30Rev+p
3IK27IK22GPDp
4IK27IK22GPDp
5IK27IK22GPDp
6IK21IK22GPDp
7IK21IK22GPDp
8IK21IK22GPDp

PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 15sec
60.0°C : 15sec
72.0°C : 30sec
72.0°C : 2min
4.0°C : Infinate
X35 cycle

09/04

PCR

 Fukuda
lanePrimer FowardPrimer ReveseTemplate
1eef-1g Feef-1gR3
2eef-1g Feef-1gR4
3AK1FAK1R5
4AK1FAK1R6
5asbFasbR7
6asbFasbR8

PCR Polymerase : KAPA Fast
95.0°C : 3min
95.0°C : 15sec
60.0°C : 15sec
72.0°C : 12sec
72.0°C : 48sec
4.0°C : Infinate
X35 cycle

09/05

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1M13FM13R161
2M13FM13R161
3M13FM13R162
4M13FM13R163
5M13FM13R166
6M13FM13R167
7M13FM13R168
8M13FM13R142
9IK60M13R161
10IK60M13R161
11M13FIK60162
12IK60M13R163
13IK60M13R166
14M13FIK60167
15M13FIK60168
16M13FIK60142

PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 20sec
61.0°C : 20sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/05

Restriction Enzyme digestion

 Yoshimto
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)RE2 nameRE2 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
1gfpp5.0EcoRI2.0NoneNoneH1.010.0
2gfpp5.0Not11.0HindⅢ1.0K1.010.0

09/05

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK21IK28Rev, p, t
2IK21IK28Rev+p, t
3IK27IK28Rev, p, t
4IK27IK28Rev+p, t
5IK21IK24Rev, p, t
6IK21IK24Rev+p, t

PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 15sec
55.0°C : 15sec
72.0°C : 30sec
72.0°C : 2min
4.0°C : Infinate
X35 cycle

09/05

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK21IK309/5 PCR p Rev,pro
2IK21IK309/5 PCR p Rev,pro
3IK21IK309/5 PCR p Rev,pro
4IK21IK309/5 PCR p Rev,pro
5IK29IK249/5 PCR p Rev,ter
6IK29IK249/5 PCR p Rev,ter
7IK29IK249/5 PCR p Rev,ter
8IK29IK249/5 PCR p Rev,ter

PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 15sec
55.0°C : 15sec
72.0°C : 30sec
72.0°C : 2min
4.0°C : Infinate
X35 cycle
PCR pulification: 5.0
Gel extraction: 5.0

09/05

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK21IK22GPD1promoter
2IK21IK22GPD1promoter
3IK21IK22GPD1promoter
4IK21IK22GPD1promoter
5IK21IK22GPD1promoter

PCR Polymerase : 2*KAPA Robust
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 1min
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/05

PCR

 Fukuda
lanePrimer FowardPrimer ReveseTemplate
1M13FM13R9
2M13FM13R10
314-3-3 zeta F14-3-3 zeta R9
414-3-3 zeta F14-3-3 zeta R10
5tropomyosin Ftropomyosin R11
6tropomyosin Ftropomyosin R12

PCR Polymerase : KAPA Fast
95.0°C : 3min
95.0°C : 15sec
60.0°C : 15sec
72.0°C : 12sec
72.0°C : 48sec
4.0°C : Infinate
X35 cycle

09/05

PCR

 Fukuda
lanePrimer FowardPrimer ReveseTemplate
1tropomyosin Ftropomyosin R11
2tropomyosin Ftropomyosin R11
3tropomyosin Ftropomyosin R11
4tropomyosin Ftropomyosin R12
5tropomyosin Ftropomyosin R12
6tropomyosin Ftropomyosin R12
714-3-3 pro F14-3-3 pro R13
814-3-3 pro F14-3-3 pro R13
914-3-3 pro F14-3-3 pro R13
1014-3-3 pro F14-3-3 pro R14
1114-3-3 pro F14-3-3 pro R14
1214-3-3 pro F14-3-3 pro R14

PCR Polymerase : KAPA Fast
95.0°C : 3min
95.0°C : 15sec
60.0°C : 15sec
72.0°C : 12sec
72.0°C : 48sec
4.0°C : Infinate
X35 cycle

09/05

PCR

 Fukuda
lanePrimer FowardPrimer ReveseTemplate
1tropomyosin Ftropomyosin R11
2tropomyosin Ftropomyosin R11
3tropomyosin Ftropomyosin R11
4tropomyosin Ftropomyosin R12
5tropomyosin Ftropomyosin R12
6tropomyosin Ftropomyosin R12
7eef-1g Feef-1g R4
8AK1 FAK1 R5
9M13FM13R9
10M13FM13R9
11M13FM13R9
12M13FM13R9
13M13FM13R10
14M13FM13R10
15M13FM13R10
16M13FM13R10

PCR Polymerase : KAPA Fast
95.0°C : 3min
95.0°C : 15sec
60.0°C : 15sec
72.0°C : 12sec
72.0°C : 48sec
4.0°C : Infinate
X35 cycle

09/06

Restriction Enzyme digestion

 Yoshimoto
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)RE2 nameRE2 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
1gfpp5.0EcoRI2.0NoneNoneH1.010.0
2gfpp5.0Not11.0HindⅢ1.0K1.010.0

09/06

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1M13FM13R163-1
2M13FM13R163-2
3M13FM13R164
4M13FM13R142

PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 20sec
61.0°C : 20sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/06

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK29IK24Rev+t
2IK29IK24Rev+t
3IK29IK24Rev+t
4IK29IK24Rev+t
5IK21IK24Rev+t, pro PCRp
6IK29IK30Rev7(plasmid)
7IK29IK30Rev7(plasmid)
8IK29IK30Rev7(plasmid)

PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 15sec
55.0°C : 15sec
72.0°C : 30sec
72.0°C : 2min
4.0°C : Infinate
X35 cycle

09/06

PCR

 Dou
lanePrimer FowardPrimer ReveseTemplate
1IK21IK22P1
2IK21IK22P1
3IK21IK22P1
4IK21IK22P1

PCR Polymerase : 2X KAPA Robust
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 1min
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/07

PCR

 Dou
lanePrimer FowardPrimer ReveseTemplate
1IK21IK22p+Rev
2IK21IK22p+Rev

PCR Polymerase : 2X KAPA Robust
95.0°C : 3min
95.0°C : 15sec
55.0°C : 15sec
72.0°C : 1min
72.0°C : 1min
4.0°C : Infinate
X35 cycle

09/07

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK60M13R142 PCR
2IK60M13R142-1
3IK60M13R142-2
4IK60M13R142-3
5IK60M13R142-4
6IK60M13R142-5

PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 15sec
60.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/07

PCR

 Fukuda
lanePrimer FowardPrimer ReveseTemplate
1IK61IK62K733018
2IK61IK62K731250

PCR Polymerase : KAPA Robust
95.0°C : 3min
95.0°C : 15sec
60.0°C : 15sec
72.0°C : 15sec
72.0°C : 3min
4.0°C : Infinate
X35 cycle

09/07

PCR

 Dou
lanePrimer FowardPrimer ReveseTemplate
1IK21IK22P1
2IK21IK22P1
3IK21IK22P1

PCR Polymerase : 2X KAPA Robust
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 1min
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/08

Restriction Enzyme digestion

 Yoshimoto
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)RE2 nameRE2 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
11415.0XbaI1.0NotI0.5K1.010.0
21445.0XbaI1.0NotI0.5K1.010.0
31435.0XbaI1.0NotI0.5K1.010.0
41455.0XbaI1.0NotI0.5K1.010.0

09/08

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK23IK28Rev3
2IK23IK28Rev4
3IK23IK282(nega)
4IK23IK28Rev new
5IK29IK30Rev3
6IK29IK30Rev4
7IK29IK302(nega)
8IK29IK30Rev new

PCR Polymerase : KAPA Robust
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 1min
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/08

PCR

 Dou
lanePrimer FowardPrimer ReveseTemplate
1IK21IK22p(blue font)
2IK21IK22p(blue font)
3IK21IK22p(blue font)
4IK21IK22p(blue font)

PCR Polymerase : 2X KAPA Robust
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 1min
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/08

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK23IK2499(nega)
2IK23IK24101
3IK23IK24Rev3
4IK23IK24Rev4
5IK23IK24Rev5
6IK23IK24Rev6
7IK23IK24T(posi)
8IK29IK3099
9IK29IK30Rev5
10IK29IK30Rev6

PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 15sec
55.0°C : 15sec
72.0°C : 30sec
72.0°C : 2min
4.0°C : Infinate
X35 cycle

09/08

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK21IK24all
2IK21IK24all
3IK21IK22promoter
4IK21IK22promoter
5IK21IK22promoter
6IK21IK22promoter
7IK29IK30Rev
8IK29IK30Rev
9IK29IK30nega 99
10IK29IK30water
11IK29IK24Rev+t
12IK29IK24Rev+t
13IK23IK24plasmid
14IK23IK24nega 99
15IK23IK24posi(T PCRp)
16IK23IK24posi(T PCRp)
17IK23IK24posi(T PCRp)
18IK23IK24posi(T PCRp)
19IK23IK24water

PCR Polymerase : go taq
95.0°C : 2min
95.0°C : 1min
55.0°C : 1min
72.0°C : 1min
72.0°C : 5min
10.0°C : Infinate
X35 cycle

09/08

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK21IK22PCRp X10
2IK21IK22PCRp X10
3IK21IK22genome X10
4IK21IK22genome
5IK21IK22water
6IK21IK24all
7IK21IK24p(PCRp),Rev+t gel ex
8IK21IK248/20 PCRp,Rev+t gel ex
9IK21IK24RA IK21X10
10IK21IK24RA
11IK21IK24water

PCR Polymerase : go taq
95.0°C : 2min
95.0°C : 1min
55.0°C : 1min
72.0°C : 2min
72.0°C : 5min
10.0°C : Infinate
X35 cycle

09/09

Ligation

 Yoshimoto
sampleVector nameVector amount(ul)Insert nameInsert amount(ul)
11432141.02.0
21452144.02.0
31432144.02.0

09/09

Restriction Enzyme digestion

 Yoshimoto
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)RE2 nameRE2 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
114220.0XbaI4.0NotI2.0K4.040.0

09/09

Ligation

 Yoshimoto
sampleVector nameVector amount(ul)Insert nameInsert amount(ul)
13230.34GFPp(gal1)12.5

09/10

Restriction Enzyme digestion

 Yoshimoto
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)RE2 nameRE2 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
1142-35.0XbaI0.5NotI0.5K1.010.0
2142-55.0XbaI0.5NotI0.5K1.010.0
3161-15.0XbaI0.5NotI0.5K1.010.0
4163-15.0XbaI0.5NotI0.5K1.010.0
51645.0XbaI0.5NotI0.5K1.010.0
61665.0XbaI0.5NotI0.5K1.010.0

09/14

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1M13FIK60162
2IK29IK24Rev
3IK40IK55GFP

PCR Polymerase : go taq
95.0°C : 2min
95.0°C : 1min
58.0°C : 1min
72.0°C : 1min
72.0°C : 2min
10.0°C : Infinate
X35 cycle

09/14

Ligation

 Yoshimoto
sampleVector nameVector amount(ul)Insert nameInsert amount(ul)
1YEplac195/sma10.34163.013.1
2YEplac195/sma10.34166.017.25
31432141.02.0
41452144.02.0
51432144.02.0

09/14

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK21IK22genome
2IK21IK22genome
3IK121IK122genome
4IK121IK122genome
5IK129IK130Rev
6IK129IK130Rev
7IK129IK130nega 99
8IK129IK130water
9IK21IK22water
10IK121IK122water

PCR Polymerase : 2X primestar
98.0°C : 3min
98.0°C : 10sec
55.0°C : 15sec
72.0°C : 5sec
72.0°C : 3min
4.0°C : Infinate
X35 cycle
PCR pulification: 1,2
PCR pulification: 4,5
Gel extraction: 1,2
Gel extraction: 4,5

09/14

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK121IK122p
2IK121IK122p
3IK21IK22p
4IK21IK22p
5IK121IK122genome
6IK121IK122genome
7IK21IK24p1, Rev+t
8IK21IK24p1, Rev+t
9IK129IK24Rev new, t
10IK21IK130p, Rev new
11IK129IK24water
12IK21IK130water

PCR Polymerase : prime star max
98.0°C : 3min
98.0°C : 10sec
55.0°C : 15sec
72.0°C : 5sec
72.0°C : 3min
4.0°C : Infinate
X35 cycle

09/15

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK129IK130Rev
2IK129IK130Rev
3IK129IK130Rev
4IK21IK24OP2+(Rev+t)
5IK129IK24NR1+t
6IK21IK130NR1+OP2

PCR Polymerase : primestar
98.0°C : 3min
98.0°C : 10sec
55.0°C : 5sec
72.0°C : 1min30sec
72.0°C : 3min
4.0°C : Infinate
X35 cycle

09/15

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK129IK24NR1+t
2IK129IK24NR1+t
3IK129IK24NR1+t
4IK129IK24NR1+t
5IK129IK24water
6K129IK130rev
7K129IK130rev

PCR Polymerase : primestar
98.0°C : 3min
98.0°C : 10sec
55.0°C : 5sec
72.0°C : 1min30sec
72.0°C : 3min
4.0°C : Infinate
X35 cycle
PCR pulification: 1~5
PCR pulification: 6,7
Gel extraction: 1~5
Gel extraction: 6,7

09/15

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK129IK24RT1
2IK121IK122genome
3IK121IK122genome
4IK121IK122genome
5IK21IK24RT1+OP1
6IK21IK24RT1+OP2
7IK121IK24RT1+NP1

PCR Polymerase : primestar
98.0°C : 3min
98.0°C : 10sec
55.0°C : 5sec
72.0°C : 1min30sec
72.0°C : 3min
4.0°C : Infinate
X35 cycle

09/15

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK121IK24RT1+NP1
2IK121IK24RT1+NP1
3IK121IK24RT1+NP1
4IK121IK24RT1+NP1
5IK129IK24RT1
6IK121IK24water

PCR Polymerase : primestar
98.0°C : 3min
98.0°C : 10sec
55.0°C : 5sec
72.0°C : 1min30sec
72.0°C : 3min
4.0°C : Infinate
X35 cycle
PCR pulification: 1~4
Gel extraction: 1~4

09/16

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK60M13R161-1
2IK60M13R161-2
3IK60M13R163-1
4IK60M13R163-2
5IK60M13R163-3
6IK60M13R163-4
7IK60M13R166-1
8IK60M13R166-2
9IK60M13R166-3
10IK60M13R166-4
11IK60M13R166-5
12IK60M13R166-6
13IK60M13R162-1
14IK60M13R162-3
15IK60M13R161-1
16IK60M13R165-2
17IK60M13R165-3
18IK60M13R165-4
19IK60M13R165-5
20IK60M13R167-1
21IK60M13R167-2
22IK60M13R167-3
23M13RIK40Gal1GFP 1
24M13RIK40Gal1GFP 2
25M13RIK40Gal1GFP 3
26M13RIK40Gal1GFP 4

PCR Polymerase : go taq
95.0°C : 2min
95.0°C : 1min
60.0°C : 1min
72.0°C : 2min
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/16

Ligation

 Ohara
sampleVector nameVector amount(ul)Insert nameInsert amount(ul)
13250.528prt20.26
23253.65prt27.33

09/16

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK121IK24prt2
2IK121IK24prt3
3IK121IK24prt3
4IK121IK24prt3
5IK121IK24prt3
6IK121IK122genome

PCR Polymerase : primestar
98.0°C : 3min
98.0°C : 10sec
55.0°C : 5sec
72.0°C : 10sec
72.0°C : 2min
4.0°C : Infinate
X35 cycle
PCR pulification: 2~5
Gel extraction: 2~5

09/16

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK60M13R142-5
2IK60M13R142-5
3M13FIK60143-1
4M13FIK60143-2
5M13FIK60143-3
6M13FIK60145-1
7M13FIK60145-2
8M13FIK60145-3

PCR Polymerase : go taq
95.0°C : 2min
95.0°C : 1min
60.0°C : 1min
72.0°C : 2min
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/16

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1M13FM13Rgenome

PCR Polymerase : go taq
95.0°C : 2min
95.0°C : 1min
60.0°C : 1min
72.0°C : 2min
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/16

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK60M13F164-1
2IK60M13F164-2
3M13FIK40Gal1 GFP 1
4M13FIK40Gal1 GFP 2
5M13FIK40Gal1 GFP 3
6M13FIK40Gal1 GFP 4

PCR Polymerase : go taq
95.0°C : 2min
95.0°C : 1min
60.0°C : 1min
72.0°C : 1min
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/17

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK121'IK24N(blue colony)
2IK121'IK241(Rev white)
3IK121'IK242
4IK121'IK243
5IK121'IK24prt3
6IK121'IK122'genome

PCR Polymerase : prime star
98.0°C : 3min
98.0°C : 10sec
55.0°C : 5sec
72.0°C : 10sec
72.0°C : 2min
4.0°C : Infinate
X35 cycle

09/17

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1M13FM13R172-0
2M13FM13R172-1
3M13FM13R172-2
4M13FM13R172-3
5M13FM13R172-4
6M13FM13R172-5
7M13FM13R172-6
8M13FM13R172-8
9M13FM13R172-9
10M13FM13R172-10
11M13FM13R172-11
12M13FM13R172-12
13M13FM13R172-13
14M13FM13R114-11
15M13FM13R114-13
16M13FM13R114-0
17M13FM13R171-1-1
18M13FM13R171-2
19M13FM13R174-1
20M13FM13R175-1
21M13FM13R175-2
22M13FM13R175-3
23M13FM13R175-4
24M13FM13R176-0
25M13FM13R176-1
26M13FM13R176-2
27M13FM13R177-1
28M13FM13R177-2
29M13FM13R177-3
30M13FM13R177-4
31M13FM13R177-5
32M13FM13Rwater
33M13FM13R26(GPD AK1)

PCR Polymerase : go taq
95.0°C : 2min15sec
95.0°C : 1min
55.0°C : 1min
72.0°C : 2min
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/17

Ligation

 Ohara
sampleVector nameVector amount(ul)Insert nameInsert amount(ul)
13250.528prt411.7

09/18

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK40IK47114-0
2IK55IK40171-1
3IK40IK41172-1
4IK60AK1F176-0
5IK60AK1F177-1
6IK60AK1F177-2

PCR Polymerase : go taq
95.0°C : 2min15sec
95.0°C : 1min
58.0°C : 1min
72.0°C : 50sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/18

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK60M13R104-1
2IK60M13R104-2

PCR Polymerase : go taq
95.0°C : 2min15sec
95.0°C : 1min
60.0°C : 1min
72.0°C : 5min
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/19

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1PrimerFPrimerR162-1
2PrimerFPrimerR162-1
3PrimerFPrimerR162-1
4PrimerFPrimerR162-1
5PrimerFPrimerRRev
6PrimerFPrimerRRev
7PrimerFPrimerRRev
8PrimerFPrimerRRev

PCR Polymerase : go taq
95.0°C : 2min15sec
95.0°C : 1min
55.0°C : 1min
72.0°C : 2min15sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/19

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1Ik29IK30OR1-1
2Ik29IK30OR1-2
3Ik29IK30OR1-3
4Ik29IK30OR1-4
5Ik29IK30OR1 nega
6Ik29IK30R2

PCR Polymerase : go taq
95.0°C : 2min15sec
95.0°C : 1min
55.0°C : 1min
72.0°C : 2min15sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/19

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK60M13R162-1
2IK60M13R162-2
3IK60M13R162-3

PCR Polymerase : go taq
95.0°C : 2min15sec
95.0°C : 1min
55.0°C : 1min
72.0°C : 2min15sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/19

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK89IK90EGFP
2IK89IK90EGFP
3IK84IK85REV
4IK86IK87REV

PCR Polymerase : go taq
95.0°C : 2min15sec
95.0°C : 1min
55.0°C : 1min
72.0°C : 2min15sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
PCR pulification: lane1,2
PCR pulification: lane3
Gel extraction: lane1,2
Gel extraction: lane3

09/20

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK40IK55171
2IK40IK55171
3IK40IK55171
4IK40IK55171
5IK40IK55171
6IK40IK55171
7IK40IK60177
8IK40IK60177
9IK40IK41172
10IK40IK41172
11IK40IK41172
12IK40IK41172
13IK40IK41172
14IK40IK41172
15IK40IK60176
16IK40IK60176
17IK46M13F148
18IK46M13F148
19IK46M13F148
20IK46M13F148
21IK46M13F148

PCR Polymerase : go taq
95.0°C : 2min15sec
95.0°C : 1min
55.0°C : 1min
72.0°C : 2min15sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/21

PCR

 Yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1M13FM13ROR-1(REV plasmid)
2M13FM13ROR-2
3M13FM13ROR-3
4M13FM13ROR-4
5M13FM13ROR-5
6IK23Ik24OR-1
7IK23Ik24OR-2
8IK23Ik24OR-3
9IK23Ik24OR-4
10IK23Ik24OR-5

PCR Polymerase : go taq
95.0°C : 2min15sec
95.0°C : 1min
55.0°C : 1min
72.0°C : 2min15sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

09/22

PCR

 Ohara
lanePrimer FowardPrimer ReveseTemplate
1IK121IK24prt3
2IK121IK24num99
3IK121IK24OR1-1
4IK121IK24OR1-2
5IK121IK24OR1-3
6IK121IK24OR1-5
7IK121IK24OR1-4

PCR Polymerase : prime star max
98.0°C : 3min
98.0°C : 10sec
55.0°C : 5sec
72.0°C : 10sec
72.0°C : 2min
4.0°C : Infinate
X35 cycle

09/22

PCR

 Dou
lanePrimer FowardPrimer ReveseTemplate
1IK89IK90EGFP2-1
2IK89IK90EGFP2-2

PCR Polymerase : go taq
95.0°C : 2min15sec
95.0°C : 1min
55.0°C : 1min
72.0°C : 2min15sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

10/04

Restriction Enzyme digestion

 yoshimoto
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)RE2 nameRE2 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
1Template 55.0Xba0.025EcoR10.025M1.010.0
2Template 55.0Not10.025EcoR10.025H1.010.0
3Template 55.0Not10.025Xba10.025H1.010.0

10/04

PCR

 ohara
lanePrimer FowardPrimer ReveseTemplate
1OD21OD23eGFP plasmid
2OD21OD23eGFP plasmid
3OD22OD23eGFP plasmid
4OD22OD23eGFP plasmid
5OD31OD32T7-RRE c
6OD31OD32T7-RRE c

PCR Polymerase : prime star max
98.0°C : 10sec
98.0°C : 10sec
55.0°C : 5sec
72.0°C : 10sec
72.0°C : 10sec
15.0°C : Infinate
X35 cycle

10/04

Restriction Enzyme digestion

 Yoshimoto
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)RE2 nameRE2 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
1pSB1C315.0Xba15.0Spe15.0M12.0100.0
2pSB1C315.0Xba15.0Pst15.0M12.0100.0

10/05

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1vf vrColony1~20(191)
2vf vrColony21~40(192)
3vf vrColony41~60(193)
4vf vrColony61~80(194)

PCR Polymerase : prime star max
98.0°C : 10sec
98.0°C : 10sec
55.0°C : 5sec
72.0°C : 10sec
72.0°C : 10sec
15.0°C : Infinate
X35 cycle

10/05

Restriction Enzyme digestion

 yoshimoto
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
1template5.0Pst10.0510xH1.010.0

10/06

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1OD41OD42Gal-GFP
2OD41OD43Gal-GFP
3OD41OD44Gal-GFP
4IK61OD52Gal-GFP
5OD53OD54Gal-GFP

PCR Polymerase : prime star max
98.0°C : 10sec
98.0°C : 10sec
55.0°C : 5sec
72.0°C : 10sec
72.0°C : 10sec
15.0°C : Infinate
X35 cycle

10/07

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK61OD54T7-RRE
2IK61OD54fukuda GFP

PCR Polymerase : pfu
95.0°C : 2min
95.0°C : 1min
55.0°C : 30sec
72.0°C : 4min
72.0°C : 4min
15.0°C : Infinate
X40 cycle

10/07

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1OD11OD12GPD-gfp
2OD11OD13GPD-gfp
3OD41OD42Gal-GFP
4OD41OD43Gal-GFP
5OD41OD44Gal-GFP
6OD31OD32T7-RRE
7IK61IK54fukuda GFP + T7RRE
8IK61IK54fukuda GFP + T7RRE

PCR Polymerase : pfu
95.0°C : 2min
95.0°C : 1min
55.0°C : 30sec
72.0°C : 4min
72.0°C : 4min
15.0°C : Infinate
X40 cycle

10/09

Restriction Enzyme digestion

 yoshimoto
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)RE2 nameRE2 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
1T7 RRE15.0Xba13.0Pst13.0M3.030.0

10/09

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK61IK54GFP+RRE

PCR Polymerase : pfu
95.0°C : 2min
95.0°C : 1min
55.0°C : 30sec
72.0°C : 4min
72.0°C : 4min
15.0°C : Infinate
X40 cycle

10/09

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1OD41OD42Gal-gfp
2OD41OD44Gal-gfp

PCR Polymerase : pfu
95.0°C : 2min
95.0°C : 1min
55.0°C : 30sec
72.0°C : 4min
72.0°C : 4min
15.0°C : Infinate
X35 cycle

10/12

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1OD61OD12TDH3-eGFP
2OD62OD12TDH3-eGFP
3OD63OD12TDH3-eGFP
4OD61OD13TDH3-eGFP
5OD62OD13TDH3-eGFP
6OD63OD13TDH3-eGFP
7OD73IK62GFP
8OD45OD42GFP
9OD45OD43GFP
10OD45OD44GFP
11IK61OD74GFP

PCR Polymerase : KAPA HiFi fast
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

10/12

Restriction Enzyme digestion

 Yoshimoto
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
1Template145.0Xba11.0M1.010.0
2Template1005.0Xba11.0M1.010.0

10/12

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1OD62OD12yeast genome
2OD62OD13yeast genome
3OD73IK62fukuda gfp
4OD45OD42gal gfp
5IK61OD74fukuda gfp

PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

10/13

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1OD62OD12yeast genome
2OD62OD12yeast genome
3OD62OD13TDH3-eGFP
4OD62OD13TDH3-eGFP
5OD73IK62fukuda GFP
6IK61OD71fukuda GFP
7OD72OD54T7-RRE

PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

10/14

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1OD62OD12yeast genome
2OD62OD13yeast genome
3OD45OD42gal1 GFP
4IK61OD74fukuda GFP
5OD73IK62fukuda GFP

PCR Polymerase : KAPA
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

10/14

PCR

 ohara
lanePrimer FowardPrimer ReveseTemplate
1IK103IK104165 gBlock
2IK107IK108161 gBlock
3IK101IK102161 gBlock
4IK105IK106163 gBlock

PCR Polymerase : yoshimoto
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

10/14

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1OD45OD42gal1 GFP
2IK61OD74fukuda GFP
3OD73IK62fukuda GFP
4IK61OD54fukuda GFP

PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

10/14

Restriction Enzyme digestion

 yoshimoto
laneSample DNA nameDNA amount (ul)RE1 nameRE1 amount(ul)Buffer nameBuffer amount(ul)Total volume(ul)
1template15.0Xba13.0M3.0100.0
2template15.0Xba15.0M3.0100.0

10/14

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1OD31OD32T7+RRE
2OD22OD23eGFP

PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

10/15

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK61OD546+7

PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

10/15

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK85IK86rev
2IK87Ik88rev

PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
PCR pulification: 1~2
Gel extraction: 1~2

10/15

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1OD31OD32T7+RRE
2OD22OD23TDH3promoter

PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

10/15

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1OD31OD32T7+RRE
2OD22OD23TDH3promoter
3OD45OD42Gal eGFP

PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X30 cycle

10/15

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK105Ik106Gblock 163

PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

10/15

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK105IK108Gal1 loop
2IK101Ik104GFP loop

PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle

10/15

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK85IK86Rev parts
2IK87IK88Rev parts
3IK61OD54T7-RRE-GFP

PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X30 cycle

10/17

PCR

 yoshitomo
lanePrimer FowardPrimer ReveseTemplate
1IK61OD544
2IK61OD545
3IK61OD547

PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X30 cycle

10/17

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK87IK88Rev
2IK105IK108Gal1-loop

PCR Polymerase :
95.0°C : 5min
98.0°C : 20sec
55.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X30 cycle

10/18

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK105IK108template1

PCR Polymerase :
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X30 cycle

10/18

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1IK107IK108Gal1pro-loop
2IK105IK106Gal1pro-loop

PCR Polymerase :
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X30 cycle

10/19

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1OD22OD23GPD-loop
2IK87IK88Gal1-loop
3IK105IK108T7-RRE
4IK101IK104TDH3-eGFP
5OD31OD32Rev

PCR Polymerase :
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X30 cycle

10/20

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1M13FM13RColony 1-10 (RRE)
2M13FM13RColony11-17 (TDH3-eGFP)
3M13FM13RColony 18-21 (Rev)
4M13FM13RColony 22-23 (GPDp-loop)
5M13FM13RColony24-28 (GPDp-loop)
6M13FM13RColony 29-32 (T7-RRE)
7M13FM13RColony 33-35 (TDH3-eGFP)
8M13FM13RColony 36-37 (GPDp-loop)
9M13FM13RColony 38-40 (Gal1-loop)
10M13FM13RColony 41-43 (Rev)
11M13FM13RColony 44 (GPDp-loop)

PCR Polymerase :
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X30 cycle

10/22

PCR

 yoshimoto
lanePrimer FowardPrimer ReveseTemplate
1OD22OD23TDH3 GFP
2IK87IK88Rev
3IK105IK108Gal1
4IK101IK104GPD1
5OD31OD32T7-RRE

PCR Polymerase :
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X30 cycle