Difference between revisions of "Team:Munich/cas13a.html"
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| Line 8: | Line 8: | ||
<td>Participants:</td> | <td>Participants:</td> | ||
| − | <td>Katja</td> | + | <td>Katja Neishsalo</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Protocol:</td> | <td>Protocol:</td> | ||
| − | <td> | + | <td> |
| + | <a href="https://static.igem.org/mediawiki/2018/7/7e/T--Munich--WL1_Restriction_Digest.pdf" target="_blank">Restriction Digest</a>, | ||
| + | <a href="https://international.neb.com/protocols/2012/12/11/gibson-assembly-protocol-e5510" target="_blank">Gibson Ligation</a>, | ||
| + | <a href="https://static.igem.org/mediawiki/2018/a/a5/T--Munich--WL1_Chemical_transformation.pdf" target="_blank">Chemical Transformation</a> | ||
| + | </td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| Line 21: | Line 25: | ||
<tr> | <tr> | ||
<td>Results:</td> | <td>Results:</td> | ||
| − | <td> | + | <td>Restriction digest showed false results; Thomas (supervisor) and Katja decided to redo the experiment</td> |
</tr> | </tr> | ||
</table> | </table> | ||
| Line 31: | Line 35: | ||
<td>Participants:</td> | <td>Participants:</td> | ||
| − | <td>Katja</td> | + | <td>Katja Neishsalo</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Protocol:</td> | <td>Protocol:</td> | ||
| − | <td>Gibson | + | <td> |
| + | <a href="https://static.igem.org/mediawiki/2018/7/7e/T--Munich--WL1_Restriction_Digest.pdf" target="_blank">Restriction Digest</a>, | ||
| + | <a href="https://international.neb.com/protocols/2012/12/11/gibson-assembly-protocol-e5510" target="_blank">Gibson Ligation</a>, | ||
| + | <a href="https://international.neb.com/protocols/2015/11/20/monarch-plasmid-dna-miniprep-kit-protocol-t1010" target="_blank">Miniprep</a>, | ||
| + | <a href="https://static.igem.org/mediawiki/2018/a/a5/T--Munich--WL1_Chemical_transformation.pdf" target="_blank">Chemical Transformation</a> | ||
| + | </td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| Line 44: | Line 53: | ||
<tr> | <tr> | ||
<td>Results:</td> | <td>Results:</td> | ||
| − | <td> | + | <td>Some colonies. Sample DNA was prepped and digested to verify the plasmid. Restriction digest gave no correct bands for the plasmid. We suspected that the Cas13a template is not good anymore.</td> |
</tr> | </tr> | ||
</table> | </table> | ||
| − | <h4>Test | + | <h4>Test Digest of p2ct_his_MBP_LBU_c2c2_WT</h4> |
<em>2018/05/28</em> | <em>2018/05/28</em> | ||
<table class="table table-borderless"> | <table class="table table-borderless"> | ||
| Line 54: | Line 63: | ||
<td>Participants:</td> | <td>Participants:</td> | ||
| − | <td>Katja</td> | + | <td>Katja Neishsalo</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Protocol:</td> | <td>Protocol:</td> | ||
| − | <td> | + | <td> |
| + | <a href="https://static.igem.org/mediawiki/2018/7/7e/T--Munich--WL1_Restriction_Digest.pdf" target="_blank">Restriction Digest</a> | ||
| + | </td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Notes:</td> | <td>Notes:</td> | ||
| − | <td>SalI NheI | + | <td>SalI NheI |
</td> | </td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Results:</td> | <td>Results:</td> | ||
| − | <td> | + | <td>Bands were as expected <figure class="figure"> |
| + | <img src="https://static.igem.org/mediawiki/2018/5/50/T--munich--Cas13a-testdigestions180528.jpg " class="figure-img img-fluid rounded" alt=" "> | ||
| + | <figcaption class="figure-caption"></figcaption> | ||
| + | </figure> | ||
| + | </td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
Latest revision as of 00:44, 18 October 2018
Assembling p2ct_his_MBP_LBU_c2c2_gRNA-EndoA
2018/04/24 - 2018/04/26| Participants: | Katja Neishsalo |
| Protocol: | Restriction Digest, Gibson Ligation, Chemical Transformation |
| Notes: | Digest of gRNA-EndoA gBlock with NheI-HF and consequent heat inactivation 80°C. Samples were gibson ligated into p2ct_his_MBP_LBU_c2c2 and E.Coli NEB Turbo was transformed. Sample DNA was preped and digested to verify the correctness. |
| Results: | Restriction digest showed false results; Thomas (supervisor) and Katja decided to redo the experiment |
Assembling p2ct_his_MBP_LBU_c2c2_gRNA-EndoA
2018/05/16| Participants: | Katja Neishsalo |
| Protocol: | Restriction Digest, Gibson Ligation, Miniprep, Chemical Transformation |
| Notes: | 2x chemical transformation (turbo, dh5a) 1x electroporation (dh5a) |
| Results: | Some colonies. Sample DNA was prepped and digested to verify the plasmid. Restriction digest gave no correct bands for the plasmid. We suspected that the Cas13a template is not good anymore. |
Test Digest of p2ct_his_MBP_LBU_c2c2_WT
2018/05/28| Participants: | Katja Neishsalo |
| Protocol: | Restriction Digest |
| Notes: | SalI NheI |
| Results: | Bands were as expected 
|