Difference between revisions of "Team:BOKU-Vienna/Notebookdata"

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     <ul><li>Christina</li></ul>
 
     <ul><li>Christina</li></ul>
 
   </ul>   
 
   </ul>   
 +
</div>
 +
 +
<div id="date-7-10">
 +
  <h1>Monday, July 10th.</h1>
 +
  <ul>
 +
    <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>Magdalena
 +
      </ul>
 +
    <li>PCR Gel Preparation</li>
 +
      <ul>Mathias
 +
      </ul>
 +
  </ul>
 +
 +
<div id="date-7-11">
 +
  <h1>Tuesday, July 11th.</h1>
 +
  <ul>
 +
    <li>Planning & Fundraising</li>
 +
    <li>Golden Gate Assembly:</li>
 +
      <ul>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
 +
      </ul>
 +
    <li>Dilution of several primer</li>
 +
      <ul><table class="table">
 +
<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>
 +
</tbody>
 +
</table>
 +
      </ul>
 +
  </ul>
 +
 
</div>
 
</div>
  
 
</body>
 
</body>
 
</html>
 
</html>

Revision as of 11:52, 24 July 2017

Monday, July 3rd.

  • Team discussion and overview:
    • Christina, Felicia, Josef, Lisa, Magdalena, Mathias, Melanie, Michael, Mina, Rebecca, Valentin, Julian
  • Planning of the procedures including primer design
  • Preparing the growth media and autoclaving
    • LB medium preparation (4L): Mina, Michael
    • Autoclaving: Mina, Rebecca

Tuesday, July 4th.

  • Planning and fundraising:
    • Christina, Felicia, Josef, Lisa, Martin, Mathias, Melanie, Michael, Mina, Rebecca, Valentin, Julian
  • Preparation of growth media:
    • 20 plates LB medium + chloramphenicol: Mina, Lisa, Mathias
    • 2 flasks LB agar: Mina, Lisa, Mathias
  • Making cells from straind DH10B chemically competent as a control for the InterLab Study

Wednesday, July 5th.

  • Planning and fundraising:
    • Christina, Felicia, Josef, Lisa, Magdalena, Martin, Mathias, Melanie, Michael, Mina, Rebecca, Julian
  • Aliquoting 1mL LB into Eppendorf tubes
    • Lisa, Josef
  • Transferring Backbones onto agar plates:
    • Julian
  • Interlab challenge - transformation: According to protocol:
      • Lisa, Mathias, Michael, Mina, Rebecca
    • 8 plasmids plated onto agar medium: Positive control (50µL, 200µL), Negative Control (50µL, 200µL), Transformant (20µL, 100µL, 200µL), Test device 1-6 (50µL, 200µL)
    • stored at 35°C

Thursday, July 6th.

  • Team discussion, planning and fundraising
  • Counting colonies of Transformants for the Interlab Challenge:
    • Colonies 200µL: 77, 20µL: 26, 100µL: 43
    Transformants on plate Transformants on plate Transformants on plate
  • Miniprep of 8 plasmids for Golden Gate Standard - according to protocol
    • Magdalena (1), Melanie (2), Valentin (3), Rebeccca (4), Christina (5), Martin (6), Mina (7), Lisa (8)
    • Dilution of plasmids for Golden Gate: Lisa
Dilution sheet for Golden Gate Assembly

Sample Volume of AD - RO Water [µL] Volume of plasmid/fragment [µL]
BB1-C-23-Spy-Cas9(NLS)-49 71,6 28,4
BB1-T-34-sce-cyc-50 83,0 17,0
BB1-P-12-syr-pNS-J23101-25 85,1 14,9
BB1-P-12-sce-tdh3-45 82,7 17,3
BB1-T-34-eco-B1001-29 80,4 19,6
BB1-P-12-syn-pm-J23105-23 83,9 16,1
BB1-C-23-syn-GFP-30 82,9 17,1
BB1-P-12-sce-tef1-46 78,0 22,0

Friday, July 7th.

  • Team discussion, Planning and Fundraising
  • Interlab Challenge: Fluorescein fluorescence standard curve
    • Christina

Monday, July 10th.

  • InterLab Study:
    • Christina, Felicia
  • Dilution of several primers 1:10 (AD-RO Water µL45 , Primer µL5)
      • Magdalena
  • PCR Gel Preparation
      • Mathias

Tuesday, July 11th.

  • Planning & Fundraising
  • Golden Gate Assembly:
      • 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
  • Dilution of several primer
      • Abbreviation Primer
      1_fw pBAD_fw_FS_1
      2_fw RT_fw_FS2_BpiI
      2_rev RT_rev_RBS_FS_C_BpiI
      3_fw FLPe_fw_FS_C_BpiI
      3_rev FLPe_rev_FS_3_BpiI
      4_fw RepA101ts_fw_FS_23_BsaI
      4_rev RepA101ts_rev_FS_3_BsaI
      5_fw AmpR_Intron_Fw_ohne_FRT_FS_2_BsaI
      5_rev AmpR_Intron_rev_ohne_FRT_FS_3_BsaI
      6_fw KanR_CripsR_fw_FS_B_BsaI
      6_rev KanR_CrispR_rev_Fs_E_BsaI