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| Line 242: |
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| | <p></p> | | <p></p> |
| | | | |
| − | <div>
| |
| − | <header>
| |
| − | <h3>July 2017</h3>
| |
| − | </header>
| |
| − | </div>
| |
| − | <div>
| |
| − | <table>
| |
| − | <tr>
| |
| − | <th>S</th>
| |
| − | <th>M</th>
| |
| − | <th>T</th>
| |
| − | <th>W</th>
| |
| − | <th>Th</th>
| |
| − | <th>F</th>
| |
| − | <th>S</th>
| |
| − | </tr>
| |
| | | | |
| − | <tr>
| |
| − | <td></td>
| |
| − | <td></td>
| |
| − | <td></td>
| |
| − | <td></td>
| |
| − | <td></td>
| |
| − | <td></td>
| |
| − | <td>1</td>
| |
| − | </tr>
| |
| − |
| |
| − | <tr>
| |
| − | <td>2</td>
| |
| − | <td><a class="page-scroll" href="#703">3</a></td>
| |
| − | <td><a href="#704">4</a></td>
| |
| − | <td><a href="#705">5</a></td>
| |
| − | <td><a href="#706">6</a></td>
| |
| − | <td><a href="#707">7</a></td>
| |
| − | <td>8</td>
| |
| − | </tr>
| |
| − |
| |
| − | <tr>
| |
| − | <td>9</td>
| |
| − | <td><a href="#710">10</a></td>
| |
| − | <td><a href="#711">11</a></td>
| |
| − | <td><a href="#712">12</a></td>
| |
| − | <td><a href="#713">13</a></td>
| |
| − | <td><a href="#714">14</a></td>
| |
| − | <td>15</td>
| |
| − | </tr>
| |
| − |
| |
| − | <tr>
| |
| − | <td>16</td>
| |
| − | <td><a href="#717">17</a></td>
| |
| − | <td><a href="#718">18</a></td>
| |
| − | <td><a href="#719">19</a></td>
| |
| − | <td><a href="#720">20</a></td>
| |
| − | <td><a href="#721">21</a></td>
| |
| − | <td>22</td>
| |
| − | </tr>
| |
| − |
| |
| − | <tr>
| |
| − | <td>23</td>
| |
| − | <td><a href="#724">24</a></td>
| |
| − | <td><a href="#725">25</a></td>
| |
| − | <td><a href="#726">26</a></td>
| |
| − | <td><a href="#727">27</a></td>
| |
| − | <td><a href="#728">28</a></td>
| |
| − | <td>29</td>
| |
| − | </tr>
| |
| − |
| |
| − | <tr>
| |
| − | <td>30</td>
| |
| − | <td><a href="#731">31</a></td>
| |
| − | <td></td>
| |
| − | <td></td>
| |
| − | <td></td>
| |
| − | <td></td>
| |
| − | <td></td>
| |
| − | </tr>
| |
| − |
| |
| − | </table>
| |
| − | </div>
| |
| − |
| |
| − | <div id="mydiv">
| |
| − | <a class="anchor" name="703"></a>
| |
| − | <h4>Monday, July 3rd.</h4>
| |
| − | <ul class="mylist">
| |
| − | <li>Team discussion and overview:</li>
| |
| − | <ul>
| |
| − | <li>Christina, Felicia, Josef, Lisa, Magdalena, Mathias, Melanie, Michael, Mina, Rebecca, Valentin, Julian</li>
| |
| − | </ul>
| |
| − | <li>Planning of the procedures including primer design</li>
| |
| − | <li><a href="https://2017.igem.org/Team:BOKU-Vienna/Protocol#lbmedium1" target="_blank">Preparing the growth media and autoclaving</a></li>
| |
| − | <ul><li>LB medium preparation (4L): Mina, Michael</li></ul>
| |
| − | <ul><li>Autoclaving: Mina, Rebecca</li></ul>
| |
| − | </ul>
| |
| − | </div>
| |
| − |
| |
| − | <p></p>
| |
| − | <p></p>
| |
| − | <div id="mydiv">
| |
| − | <a name="704" class="anchor"></a>
| |
| − | <h4>Tuesday, July 4th.</h4>
| |
| − | <ul class="mylist">
| |
| − | <li>Planning and fundraising:</li>
| |
| − | <ul>
| |
| − | <li>Christina, Felicia, Josef, Lisa, Martin, Mathias, Melanie, Michael, Mina, Rebecca, Valentin, Julian</li>
| |
| − | </ul>
| |
| − | <li>Preparation of growth media:</li>
| |
| − | <ul><li>20 plates <a href="https://2017.igem.org/Team:BOKU-Vienna/Protocol#lbmedium1" target="_blank">LB medium</a> + chloramphenicol: Mina, Lisa, Mathias</li></ul>
| |
| − | <ul><li>2 flasks <a href="https://2017.igem.org/Team:BOKU-Vienna/Protocol#lbagar" target="_blank">LB agar</a>: Mina, Lisa, Mathias</li></ul>
| |
| − | <li>Making cells from straind DH10B chemically competent</li>
| |
| − | </ul>
| |
| − | </div>
| |
| − |
| |
| − | <p></p>
| |
| − | <p></p>
| |
| − | <div id="mydiv">
| |
| − | <a name="705" class="anchor"></a>
| |
| − | <h4>Wednesday, July 5th.</h4>
| |
| − | <ul class="mylist">
| |
| − | <li>Planning and fundraising:</li>
| |
| − | <ul>
| |
| − | <li>Christina, Felicia, Josef, Lisa, Magdalena, Martin, Mathias, Melanie, Michael, Mina, Rebecca, Julian</li>
| |
| − | </ul>
| |
| − | <li>Aliquoting 1mL LB into Eppendorf tubes</li>
| |
| − | <ul><li>Lisa, Josef</li></ul>
| |
| − | <li>Transferring Backbones onto agar plates:</li>
| |
| − | <ul><li>Julian</li></ul>
| |
| − | <li>Interlab challenge - transformation: According to <a href="https://2017.igem.org/Team:BOKU-Vienna/Protocol#transformation" target="_blank">protocol</a>:</li>
| |
| − | <ul>Lisa, Mathias, Michael, Mina, Rebecca</ul>
| |
| − | <ul><li>8 transformations - from <a href="https://2017.igem.org/Team:BOKU-Vienna/Protocol#interlab" target="_blank">Kit Plate 6 InterLab</a></li></ul>
| |
| − | <ul><li>8 plasmids plated onto agar medium: Positive control (50µL, 200µL), Negative Control (50µL, 200µL), Transformant (20µL, 100µL, 200µL), Test device 1-6 (50µL, 200µL)</li></ul>
| |
| − | <ul><li>stored at 35°C</li></ul>
| |
| − | </ul>
| |
| − | </div>
| |
| − |
| |
| − | <p></p>
| |
| − | <p></p>
| |
| − | <div id="mydiv">
| |
| − | <a name="706" class="anchor"></a>
| |
| − | <h4>Thursday, July 6th.</h4>
| |
| − | <ul class="mylist">
| |
| − | <li>Team discussion, planning and fundraising</li>
| |
| − | <li>Counting colonies of Transformants for Determination of Transformation Efficiency of our DH10B cells:</li>
| |
| − | <ul><li>Colonies 200µL: 77, 20µL: 26, 100µL: 43</li></ul>
| |
| − | <img src="https://static.igem.org/mediawiki/2017/1/17/T--BOKU-Vienna--Notebook_countedplate1.jpg" alt="Transformants on plate" height="200">
| |
| − | <img src="https://static.igem.org/mediawiki/2017/f/f6/T--BOKU-Vienna--Notebook_countedplate2.jpg" alt="Transformants on plate" height="200">
| |
| − | <img src="https://static.igem.org/mediawiki/2017/2/2c/T--BOKU-Vienna--Notebook_countedplate3.jpg" alt="Transformants on plate" height="200">
| |
| − | <li>Miniprep of 8 plasmids for Golden Gate Standard - according to <a href="https://2017.igem.org/Team:BOKU-Vienna/Protocol#miniprep" target="_blank">protocol</a></li>
| |
| − | <ul><li>Magdalena (1), Melanie (2), Valentin (3), Rebeccca (4), Christina (5), Martin (6), Mina (7), Lisa (8)</li></ul>
| |
| − | <ul><li>Dilution of plasmids for Golden Gate: Lisa</li></ul>
| |
| − | </ul>
| |
| − |
| |
| − | <h5 id="dilution">Dilution sheet for Golden Gate Assembly</h5>
| |
| − | <p></p>
| |
| − | <div>
| |
| − | <table class="table1" >
| |
| − | <thead>
| |
| − | <tr>
| |
| − | <th>Sample</th>
| |
| − | <th>Volume of AD - RO Water [µL]</th>
| |
| − | <th>Volume of plasmid/fragment [µL]</th>
| |
| − | </tr>
| |
| − | </thead>
| |
| − | <tbody>
| |
| − | <tr>
| |
| − | <td>BB1-C-23-Spy-Cas9(NLS)-49</td>
| |
| − | <td>71,6</td>
| |
| − | <td>28,4</td>
| |
| − | </tr>
| |
| − | <tr>
| |
| − | <td>BB1-T-34-sce-cyc-50</td>
| |
| − | <td>83,0</td>
| |
| − | <td>17,0</td>
| |
| − | </tr>
| |
| − | <tr>
| |
| − | <td>BB1-P-12-syr-pNS-J23101-25</td>
| |
| − | <td>85,1</td>
| |
| − | <td>14,9</td>
| |
| − | </tr>
| |
| − | <tr>
| |
| − | <td>BB1-P-12-sce-tdh3-45</td>
| |
| − | <td>82,7</td>
| |
| − | <td>17,3</td>
| |
| − | </tr>
| |
| − | <tr>
| |
| − | <td>BB1-T-34-eco-B1001-29</td>
| |
| − | <td>80,4</td>
| |
| − | <td>19,6</td>
| |
| − | </tr>
| |
| − | <tr>
| |
| − | <td>BB1-P-12-syn-pm-J23105-23</td>
| |
| − | <td>83,9</td>
| |
| − | <td>16,1</td>
| |
| − | </tr>
| |
| − | <tr>
| |
| − | <td>BB1-C-23-syn-GFP-30</td>
| |
| − | <td>82,9</td>
| |
| − | <td>17,1</td>
| |
| − | </tr>
| |
| − | <tr>
| |
| − | <td>BB1-P-12-sce-tef1-46</td>
| |
| − | <td>78,0</td>
| |
| − | <td>22,0</td>
| |
| − | </tr>
| |
| − | </tbody>
| |
| − | </table>
| |
| − | </div>
| |
| − | </div>
| |
| − | <p></p>
| |
| − | <p></p>
| |
| − | <div id="mydiv">
| |
| − | <a name="707" class="anchor"></a>
| |
| − | <h4>Friday, July 7th.</h4>
| |
| − | <ul class="mylist">
| |
| − | <li>Team discussion, Planning and Fundraising</li>
| |
| − | <li><a href="https://2017.igem.org/Team:BOKU-Vienna/Protocol#interlab" target="_blank">Interlab Challenge</a>: Fluorescein fluorescence standard curve</li>
| |
| − | <ul><li>Christina</li></ul>
| |
| − | </ul>
| |
| − | </div>
| |
| − |
| |
| − | <div id="mydiv">
| |
| − | <a name="710" class="anchor"></a>
| |
| − | <h4>Monday, July 10th.</h4>
| |
| − | <ul class="mylist">
| |
| − | <li>InterLab Study:</li>
| |
| − | <ul>
| |
| − | <li>Christina, Felicia</li>
| |
| − | </ul>
| |
| − | <li>Dilution of several primers 1:10 (AD-RO Water µL45 , Primer µL5)</li>
| |
| − | <ul><li>Magdalena</li>
| |
| − | </ul>
| |
| − | <li>PCR Gel Preparation</li>
| |
| − | <ul><li>Mathias</li>
| |
| − | </ul>
| |
| − | </ul>
| |
| − | </div>
| |
| − |
| |
| − | <p></p>
| |
| − | <p></p>
| |
| − |
| |
| − | <div id="mydiv">
| |
| − | <a name="711" class="anchor"></a>
| |
| − | <h4>Tuesday, July 11th.</h4>
| |
| − | <ul class="mylist">
| |
| − | <li>Planning & Fundraising</li>
| |
| − | <li>Golden Gate Assembly:</li>
| |
| − | <ul><li>Flpe (BB1), GFP (BB2) → Transformation (according to protocol) → Plating onto Agar (After Transformation): Selection with Ampicillin (bb1), Selection with Kanamycin(bb2), stored at 37°C over night
| |
| − | </li></ul>
| |
| − | <li>Dilution of several primer</li>
| |
| − | <ul><table class="table1">
| |
| − | <thead>
| |
| − | <tr>
| |
| − | <th>Abbreviation</th>
| |
| − | <th>Primer</th>
| |
| − | </tr>
| |
| − | </thead>
| |
| − | <tbody>
| |
| − | <tr>
| |
| − | <td>1_fw</td>
| |
| − | <td>pBAD_fw_FS_1</td>
| |
| − |
| |
| − | </tr>
| |
| − | <tr>
| |
| − | <td>2_fw</td>
| |
| − | <td>RT_fw_FS2_BpiI</td>
| |
| − |
| |
| − | </tr>
| |
| − | <tr>
| |
| − | <td>2_rev</td>
| |
| − | <td>RT_rev_RBS_FS_C_BpiI</td>
| |
| − |
| |
| − | </tr>
| |
| − | <tr>
| |
| − | <td>3_fw</td>
| |
| − | <td>FLPe_fw_FS_C_BpiI</td>
| |
| − | </tr>
| |
| − | <tr>
| |
| − | <td>3_rev</td>
| |
| − | <td>FLPe_rev_FS_3_BpiI</td>
| |
| − | </tr>
| |
| − | <tr>
| |
| − | <td>4_fw</td>
| |
| − | <td>RepA101ts_fw_FS_23_BsaI</td>
| |
| − |
| |
| − | </tr>
| |
| − | <tr>
| |
| − | <td>4_rev</td>
| |
| − | <td>RepA101ts_rev_FS_3_BsaI</td>
| |
| − | </tr>
| |
| − | <tr>
| |
| − | <td>5_fw</td>
| |
| − | <td>AmpR_Intron_Fw_ohne_FRT_FS_2_BsaI</td>
| |
| − | </tr>
| |
| − |
| |
| − | <tr>
| |
| − | <td>5_rev</td>
| |
| − | <td>AmpR_Intron_rev_ohne_FRT_FS_3_BsaI</td>
| |
| − | </tr>
| |
| − | <tr>
| |
| − | <td>6_fw</td>
| |
| − | <td>KanR_CripsR_fw_FS_B_BsaI</td>
| |
| − |
| |
| − | </tr>
| |
| − | <tr>
| |
| − | <td>6_rev</td>
| |
| − | <td>KanR_CrispR_rev_Fs_E_BsaI</td>
| |
| − | </tr>
| |
| − | <tr>
| |
| − | <td>7_fw</td>
| |
| − | <td>AmpR_Intron_Fw_FRT_FS_2_BsaI</td>
| |
| − | </tr>
| |
| − | <tr>
| |
| − | <td>7_rev</td>
| |
| − | <td>AmpR_Intron_rev_FRT_FS_3_BsaI</td>
| |
| − | </tr>
| |
| − | </tbody>
| |
| − | </table>
| |
| − | </ul>
| |
| − | <li>PCR plasmid fragments</li>
| |
| − | <ul><li>Josef</li>
| |
| − | </ul>
| |
| − | <ul><li>fragment 1: GFP+Primer binding side, fragment 2: GFP + ½ FRT sequence, fragment 3: 1 random sequence
| |
| − | </li></ul>
| |
| − | <li>Gel electrophoresis</li>
| |
| − | </ul>
| |
| − | </div>
| |
| − |
| |
| − | <p></p>
| |
| − | <p></p>
| |
| − | <div id="mydiv">
| |
| − | <a name="712" class="anchor"></a>
| |
| − | <h4>Wednesday, July 12th.</h4>
| |
| − | <ul class="mylist">
| |
| − | <li>Gel PCR for Kanamycin casette</li>
| |
| − | <ul>
| |
| − | <li>Mina, Magda</li>
| |
| − | </ul>
| |
| − | <li>Restriction digest</li>
| |
| − | <li>Gel/PCR DNA Extraction </li>
| |
| − | <li>NEB Golden Gate Assembly Mix</li>
| |
| − |
| |
| − | <li>Miniprep</li>
| |
| − | <ul><li>Lisa</li></ul>
| |
| − | <li>InterLab Study</li>
| |
| − | <ul>
| |
| − | <li>Felicia, Christina</li>
| |
| − | </ul>
| |
| − | <li>PCR and gel electrophoresis</li>
| |
| − | <ul>
| |
| − | <li>Michael</li>
| |
| − | </ul>
| |
| − | </ul>
| |
| − | </div>
| |
| − |
| |
| − | <p></p>
| |
| − | <p></p>
| |
| − | <div id="mydiv">
| |
| − | <a name="713" class="anchor"></a>
| |
| − | <h4>Thursday, July 13th.</h4>
| |
| − | <ul class="mylist">
| |
| − | <li>Miniprep: Flpe + GFP BB2, control digest</li>
| |
| − | <ul>
| |
| − | <li>Lisa, Mina, Valentin</li>
| |
| − | </ul>
| |
| − | <li>Minis onto LB Kanamycin medium</li>
| |
| − | <ul><li>Rebecca</li></ul>
| |
| − |
| |
| − | <li>Control digest - GFP</li>
| |
| − | <ul><li>Mina, Lisa</li></ul>
| |
| − |
| |
| − | <li>Gel electrophoresis</li>
| |
| − | <ul><li>Felicia, Mathias</li></ul>
| |
| − |
| |
| − | <li>Golden Gate Assembly:</li>
| |
| − | <ul><li>Joseph, Felicia, Mina, Christina, Rebecca</li></ul>
| |
| − | </ul>
| |
| − | </div>
| |
| − |
| |
| − |
| |
| − |
| |
| − | <p></p>
| |
| − | <p></p>
| |
| − | <div id="mydiv">
| |
| − | <a name="714" class="anchor"></a>
| |
| − | <h4>Friday, July 14th.</h4>
| |
| − | <ul class="mylist">
| |
| − | <li>Transformation from GG-clones in competent cells using LB medium (with Kan or Amp)</li>
| |
| − | <ul>
| |
| − | <li>Christina, Joseph, Felicia, Melanie, Rebecca </li>
| |
| − | </ul>
| |
| − |
| |
| − | <li>Golden Gate Assembly: </li>
| |
| − | <ul><li>stored at 37°C</li></ul>
| |
| − | <ul><li>Joseph, Felicia, Mina, Christina, Rebecca, Lisa, Mathias, Valentin</li></ul>
| |
| − |
| |
| − | <li>Gel electrophoresis</li>
| |
| − | <ul><li>Felicia, Valentin</li></ul>
| |
| − | <li>Purifiying DNA fragments from elektrophoresis according to given protocol in the HiYield Gel/PCR DNA Extraction Kit</li>
| |
| − | <ul><li>Felicia, Christina, Valentin, Lisa</li></ul>
| |
| − | <li>Concentration measurement of purified DNA</li>
| |
| − | <ul><li> Julian</li></ul>
| |
| − | </ul>
| |
| − | </div>
| |
| − |
| |
| − |
| |
| − | <p></p>
| |
| − | <p></p>
| |
| − |
| |
| − | <div id="mydiv">
| |
| − | <a name="717" class="anchor"></a>
| |
| − | <h4>Monday, July 17th.</h4>
| |
| − | <ul class="mylist">
| |
| − | <li>Miniprep</li>
| |
| − | <ul>
| |
| − | <li>control digest</li>
| |
| − | </ul>
| |
| − | <ul>
| |
| − | <li>BB1_10, BB1_11, BB1_12, BB2_2, BB2_3, BB2_4, BB2_5: </li>
| |
| − | </ul>
| |
| − | <ul>
| |
| − | <li>Gel electrophoresis</li>
| |
| − | </ul>
| |
| − | <ul>
| |
| − | <li>Mathias, Mina, Magda, Melanie, Lisa, Rebecca</li>
| |
| − | </ul>
| |
| − |
| |
| − | <li>Mini: p5IM5 30°C, Kryo</li>
| |
| − | <ul><li>Rebecca</li></ul>
| |
| − | <ul><li>Control Transformation</li></ul>
| |
| − |
| |
| − | <li>PCR</li>
| |
| − | <ul><li>Golden Gate: BB2_6 (Rep A, BB2_7: Cas 9, BB2_8: RT, Flpe, BB2_8 → RT, Flpe coli, BB2_1+BB2_5) + Trafo (on LB AMP)
| |
| − | </li></ul>
| |
| − | <li>Purifiying DNA fragments from elektrophoresis according to given protocol in the HiYield Gel/PCR DNA Extraction Kit</li>
| |
| − | <li>Transformation of Golden Gate products:</li>
| |
| − | <ul><li>Michi, Julian, Christina, Felicia</li></ul>
| |
| − |
| |
| − |
| |
| − | <li>Kryo for Sequencing</li>
| |
| − | <ul class="mylist">
| |
| − | <li>Kryo: tube with Glycerine + sample</li>
| |
| − | </ul>
| |
| − | </ul>
| |
| − | </div>
| |
| − |
| |
| − |
| |
| − | <p></p>
| |
| − | <p></p>
| |
| − |
| |
| − | <div id="mydiv">
| |
| − | <a name="718" class="anchor"></a>
| |
| − | <h4>Tuesday, July 18th.</h4>
| |
| − | <ul class="mylist">
| |
| − | <li>Load gel for gel electrophoresis</li>
| |
| − | <ul>
| |
| − | <li>Melanie</li>
| |
| − | </ul>
| |
| − | <li>Media preparation</li>
| |
| − | <ul>
| |
| − | <li>Melanie, Valentin</li>
| |
| − | </ul>
| |
| − | <li>Colony PCR (11 colonies)</li>
| |
| − | <ul><li>Magdalena, Felicia, Christina, Mina</li></ul>
| |
| − |
| |
| − | <li>Preparation for Sequencing (cryo)</li>
| |
| − | <ul>
| |
| − | <li>Christina</li>
| |
| − | </ul>
| |
| − | <li>more colony PCR</li>
| |
| − | <ul>
| |
| − | <li>Felicia, Christina, Valentin, Rebecca, Lisa</li>
| |
| − | </ul>
| |
| − |
| |
| − | <li>PCR Reaction with ThermoCycler</li>
| |
| − | </ul>
| |
| − | </div>
| |
| − |
| |
| − | <p></p>
| |
| − | <p></p>
| |
| − | <div id="mydiv">
| |
| − | <a name="719" class="anchor"></a>
| |
| − | <h4>Wednesday, July 19th.</h4>
| |
| − | <ul class="mylist">
| |
| − | <li>PCR gel elektrophoresis</li>
| |
| − | <ul>
| |
| − | <li>Melanie, Christina, Felicia, Magdalena</li>
| |
| − | </ul>
| |
| − | <li>Media preparation (Agar)</li>
| |
| − | <ul>
| |
| − | <li>Lisa, Rebecca</li>
| |
| − | </ul>
| |
| − | <li>Miniprep</li>
| |
| − | <ul><li>Christina, Felicia, Valentin, Rebecca</li></ul>
| |
| − |
| |
| − | <li>Preparation for Sequencing</li>
| |
| − | <ul>
| |
| − | <li>Christina, Melanie</li>
| |
| − | </ul>
| |
| − | <li>Golden Gate</li>
| |
| − | <ul>
| |
| − | <li>Mathias, Felicia, Lisa, Rebecca</li>
| |
| − | </ul>
| |
| − |
| |
| − | <li>Transformation for Golden Gate</li>
| |
| − | <ul>
| |
| − | <li>Magdalena, Lisa, Mina, Rebecca, Mathias, Julian, Michael</li>
| |
| − | </ul>
| |
| − | </ul>
| |
| − | </div>
| |
| − |
| |
| − |
| |
| − | <p></p>
| |
| − | <p></p>
| |
| − | <div id="mydiv">
| |
| − | <a name="720" class="anchor"></a>
| |
| − | <h4>Thursday, July 20th.</h4>
| |
| − | <ul class="mylist">
| |
| − | <li>Colony PCR:</li>
| |
| − | <ul><li>
| |
| − | 9 samples: BB2_1 A, B, C, D, E, F und BB2_11 A, B, C
| |
| − | </li>
| |
| − | </ul>
| |
| − | <ul>
| |
| − | <li>Lisa, Christina, Felicia, Rebecca, Lisa, Mina</li>
| |
| − | </ul>
| |
| − | <li>Golden Gate repetition (CRISPR: 5alpha, 10B):</li>
| |
| − | <ul>
| |
| − | <li>Felicia</li>
| |
| − | </ul>
| |
| − | <li>DH10B transformation of Golden Gate products (5alpha, 10B):</li>
| |
| − | <ul><li>Christina, Felicia</li></ul>
| |
| − |
| |
| − | <li>PCR #5</li>
| |
| − | <ul>
| |
| − | <li>Christina</li>
| |
| − | </ul>
| |
| − | <li>Gel electrophoresis</li>
| |
| − | <ul><li>
| |
| − | with Colony PCR products, Golden Gate products, PCR #5
| |
| − | </li></ul>
| |
| − | <ul>
| |
| − | <li>Felicia, Christina</li>
| |
| − | </ul>
| |
| − |
| |
| − | <li>DNA purification of PCR #6</li>
| |
| − | <ul>
| |
| − | <li>Christina</li>
| |
| − | </ul>
| |
| − | <li>Aliquoting Master Mix</li>
| |
| − | <ul>
| |
| − | <li>Valentin</li>
| |
| − | </ul>
| |
| − | <li>Golden Gate (GFP, BB2_16)</li>
| |
| − | <ul>
| |
| − | <li>Valentin, Mathias, Rebecca</li>
| |
| − | </ul>
| |
| − | <li>Plating repA colony onto agar</li>
| |
| − | <ul>
| |
| − | <li>Lisa, Valentin</li>
| |
| − | </ul>
| |
| − | <li>Transformation</li>
| |
| − | <ul>
| |
| − | <li>Michael, Julian, Felicia, Christina</li>
| |
| − | </ul>
| |
| − | </ul>
| |
| − | </div>
| |
| − |
| |
| − |
| |
| − |
| |
| − | <p></p>
| |
| − | <p></p>
| |
| − | <div id="mydiv">
| |
| − | <a name="721" class="anchor"></a>
| |
| − | <h4>Friday, July 21st.</h4>
| |
| − | <ul class="mylist">
| |
| − | <li>Minipreparation:</li>
| |
| − | <ul>
| |
| − | <li>BB2_11A: Christina</li>
| |
| − | </ul>
| |
| − | <li>BB3_5: Yeast in LB Hygromycin</li>
| |
| − | <li>Colony PCR</li>
| |
| − | <ul><li>Rebecca, Felicia, Melanie, Christina, Mathias</li></ul>
| |
| − | <li>Golden Gate PCR - CRISPR</li>
| |
| − | <ul><li>Christina, Magdalena, Mathias</li></ul>
| |
| − | </ul>
| |
| − | </div>
| |
| − |
| |
| − |
| |
| − | <p></p>
| |
| − | <p></p>
| |
| − | <div id="mydiv">
| |
| − | <a name="724" class="anchor"></a>
| |
| − | <h4>Monday, July 24th.</h4>
| |
| − | <ul class="mylist">
| |
| − | <li>Colony PCR: CRISPR (4x), BB3_5 (6x)</li>
| |
| − | <ul>
| |
| − | <li>Lisa, Valentin</li>
| |
| − | </ul>
| |
| − | <li>PCR: yeast sgRNA URA3, yeast sgRNA LEV2</li>
| |
| − | <ul>
| |
| − | <li>Mathias, Christina, Melanie</li>
| |
| − | </ul>
| |
| − | <li>Golden Gate</li>
| |
| − | <ul><li>BB3_2, BB3_1, BB3_3, BB3_4, BB3_6, BB2_18, BB3_8</li></ul>
| |
| − | <ul><li>Christina, Mathias</li></ul>
| |
| − | <li>Golden Gate</li>
| |
| − | <ul><li>BB1_16, BB1_17</li></ul>
| |
| − | <ul><li>Mina, Felicia</li></ul>
| |
| − | </ul>
| |
| − | </div>
| |
| − |
| |
| − | <p></p>
| |
| − | <div id="mydiv">
| |
| − | <a name="725" class="anchor"></a>
| |
| − | <h4>Tuesday, July 25th.</h4>
| |
| − | <ul class="mylist">
| |
| − | <li>Minipreparation of BB3_5B and BB3_5F:</li>
| |
| − | <ul>
| |
| − | <li>Rebecca</li>
| |
| − | </ul>
| |
| − | <li>OneTaq PCR from colonies of BB3_1; BB3_2; BB3_8 and plated onto petri dishes with LB-Hygromycine</li>
| |
| − | <ul>
| |
| − | <li> Lisa, Magdalena, Felicia, Mathias, Melanie, Christina</li>
| |
| − | </ul>
| |
| − | <li>Some agar plates (BB3_1 + 2 + 8) had too many colonies, so single colonies were plated onto new agar plates</li>
| |
| − | <ul><li>Felicia, Josef, Magda</li></ul>
| |
| − | <li>Analytical gel electrophoresis from previous OneTaq PCRs</li>
| |
| − | <ul><li>Julian, Felicia, Christina</li></ul>
| |
| − | <li>Transformation (BB1_16, BB1_17 → Hefe guide RNA)</li>
| |
| − | <ul>
| |
| − | <li> Julian, Josef</li>
| |
| − | </ul>
| |
| − | </ul>
| |
| − | </div>
| |
| − |
| |
| − | <p></p>
| |
| − | <div id="mydiv">
| |
| − | <a name="726" class="anchor"></a>
| |
| − | <h4>Wednesday, July 26th.</h4>
| |
| − | <ul class="mylist">
| |
| − | <li>Colony PCR: BB1_16, BB1_17, BB3_1 + 2 + 8 and plasmid preparation</li>
| |
| − | <ul>
| |
| − | <li>Magdalena, Lisa, Valentin, Josef</li>
| |
| − | </ul>
| |
| − | <li>Golden Gate with URA3</li>
| |
| − | <ul>
| |
| − | <li>Josef, Julian, Christina, Rebecca</li>
| |
| − | </ul>
| |
| − | <li>Miniprep: </li>
| |
| − | <ul><li>BB2_18A; BB2_18D; BB2_18F: Melanie, Felicia</li></ul>
| |
| − | <ul><li>BB2_18A; BB2_18D; BB2_18F: Valentin, Mathias</li></ul>
| |
| − | <li>PCR: #38, #39, #40</li>
| |
| − | <li>Gel/PCR DNA Extraction:</li>
| |
| − | <ul>
| |
| − | <li> Felicia</li>
| |
| − | </ul>
| |
| − | <li>OD Measurement of preculture from SC288:</li>
| |
| − | <ul>
| |
| − | <li>Mathias, Melanie</li>
| |
| − | </ul>
| |
| − | </ul>
| |
| − | </div>
| |
| − |
| |
| − | <p></p>
| |
| − | <div id="mydiv">
| |
| − | <a name="727" class="anchor"></a>
| |
| − | <h4>Thursday, July 27th.</h4>
| |
| − | <ul class="mylist">
| |
| − | <li>Colony PCR: BB3_1, 5, 2; BB2_17; BB1_18; CRISPR-PPP</li>
| |
| − | <ul>
| |
| − | <li>Magdalena, Felicia, Lisa, Mina</li>
| |
| − | </ul>
| |
| − | <li>Miniprep from BB1_16, 17</li>
| |
| − | <ul>
| |
| − | <li>Magdalena, Lisa</li>
| |
| − | </ul>
| |
| − | <li>PCR #41-43, 31-36 including Gel electrophoresis and gel DNA extraction</li>
| |
| − | <ul><li>Felicia, Rebecca, Christina, Mathias, Julian </li></ul>
| |
| − | <li>PCR: #38, #39, #40</li>
| |
| − | <li>Golden Gate</li>
| |
| − | <ul>
| |
| − | <li>BB2_10</li>
| |
| − | <li>BB2_19 and BB2_10: Josef, Felicia</li>
| |
| − | </ul>
| |
| − | <li>Preparing LB-Amp agar plates</li>
| |
| − | <ul>
| |
| − | <li>Melanie, Rebecca, Josef</li>
| |
| − | </ul>
| |
| − | </ul>
| |
| − | </div>
| |
| − |
| |
| − | <p></p>
| |
| − | <div id="mydiv">
| |
| − | <a name="728" class="anchor"></a>
| |
| − | <h4>Friday, July 28th.</h4>
| |
| − | <ul class="mylist">
| |
| − | <li>Colony PCR</li>
| |
| − | <ul><li>BB1_20 - BB1_25, BB2_23, BB2_24, BB3_8 + preculture for miniprep in Kanamycin, Hygromycin & Ampicillin Medium</li></ul>
| |
| − | <ul><li>Magdalena, Felicia, Rebecca</li></ul>
| |
| − | <ul><li>Preculture for miniprep: BB3_1, BB3_5 (Magdalena, Lisa)<li>
| |
| − |
| |
| − | </ul>
| |
| − | <li>Miniprep: CRISPR-PPP; BB3_2; BB1_18</li>
| |
| − | <ul>
| |
| − | <li>Lisa, Mina</li>
| |
| − | </ul>
| |
| − | <li>Sequencing BB3_8</li>
| |
| − | <ul><li>Christina</li></ul>
| |
| − | <li>Transformation</li>
| |
| − | <ul><li>BB2_10,19; BB1_20-25</li></ul>
| |
| − | <li>LB-Kanamycin plates + agarose gel for gel electrophoresis</li>
| |
| − | <ul>
| |
| − | <li>Valentin</li>
| |
| − | </ul>
| |
| − | <li>Gel electrophoresis</li>
| |
| − | <ul>
| |
| − | <li>CRISPR, BB1_18, BB3_2, BB2_2 and gel DNA extraction</li>
| |
| − | <li>Christina, Mathias</li>
| |
| − | <li>BB1_20, 21, 22, 23, 24, BB1_25, BB2_23, BB2_24, BB3_8</li>
| |
| − | <li>Lisa, Felicia, Rebecca</li>
| |
| − | <li>BB3_1, BB3_5</li>
| |
| − | <li>Christina</li>
| |
| − | </ul>
| |
| − | <li>Repetition of colony PCR: BB2_23 and BB3_8</li>
| |
| − | </ul>
| |
| − | </div>
| |
| − |
| |
| − | <p></p>
| |
| − | <div id="mydiv">
| |
| − | <a name="731" class="anchor"></a>
| |
| − | <h4>Monday, July 31st.</h4>
| |
| − | <ul class="mylist">
| |
| − | <li>Colony PCR</li>
| |
| − | <ul><li>BB2_10; BB2_19; BB2_20; BB3_1; BB3_6; BB3_8 and miniprep pre-cultures in Amp- or Kan- Media</li></ul>
| |
| − | <ul><li>Lisa, Magda, Mina, Rebecca, Felicia</li></ul>
| |
| − |
| |
| − | <li>Golden Gate Assembly:</li>
| |
| − | <ul>
| |
| − | <li>BB2 25-30, CRISPR 2-3 & BB3_2 + Amp, BB1_19</li>
| |
| − | </ul>
| |
| − | <ul>
| |
| − | <li> Josef, Rebecca, Lisa, Christina, Mathias</li>
| |
| − | </ul>
| |
| − | <li>Transformation of E. Coli BB2_20 +28+30, BB2_27 + 29 + CRISPR 2 & 3</li>
| |
| − | <li>Repetition of colony PCR: BB2_19 with new primers (41 fw, 42 rev)</li>
| |
| − | <ul><li>Magda, Lisa, Mina, Rebecca</li></ul>
| |
| − | <li>Gel electrophoresis</li>
| |
| − | <ul><li> Felicia, Josef</li></ul>
| |
| − | <li>Transformation of yeast</li>
| |
| − | <ul>
| |
| − | <li>Lisa, Michael, Rebecca</li>
| |
| − | </ul>
| |
| − | </ul>
| |
| − | </div>
| |
| − |
| |
| − | </body>
| |
| | | | |
| | | | |
