Difference between revisions of "Team:Munich/deletioncasette.html"
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| − | <h4>Transforming E. Coli DH5a to | + | <h4>Transforming E. Coli DH5a to Amplify pKD3 for pRED/ET Engineering</h4> |
<em>2018/05/28</em> | <em>2018/05/28</em> | ||
<table class="table table-borderless"> | <table class="table table-borderless"> | ||
| Line 13: | Line 13: | ||
<td>Protocol:</td> | <td>Protocol:</td> | ||
<td> | <td> | ||
| − | <a href="https://static.igem.org/mediawiki/2018/c/ca/T--Munich--WL1_Electrocompetent_transformation.pdf" target="_blank"> | + | <a href="https://static.igem.org/mediawiki/2018/c/ca/T--Munich--WL1_Electrocompetent_transformation.pdf" target="_blank">Electro-transformation</a> |
</td> | </td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Notes:</td> | <td>Notes:</td> | ||
| − | <td> | + | <td>No notes.</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td> | <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td> | ||
<td>No colonies, no growth; <br> | <td>No colonies, no growth; <br> | ||
| − | We decided to use a resistance cassette from pSB1C3 without FRT sites | + | We decided to use a resistance cassette from pSB1C3 without FRT sites. |
</td> | </td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
| − | <h4>Amplifying a | + | <h4>Amplifying a Selection Cassette from pSB1C3</h4> |
<em>2018/05/29</em> | <em>2018/05/29</em> | ||
<table class="table table-borderless"> | <table class="table table-borderless"> | ||
| Line 46: | Line 46: | ||
<tr> | <tr> | ||
<td>Notes:</td> | <td>Notes:</td> | ||
| − | <td> | + | <td>PCR with two primer pairs were tested to amplify a chloramphenicol resistance cassette for the knockout. Both pairs have a 50 bp overhang homology region to the knockout target in E.Coli genomes. One pair has longer annealing sequence (CAT_RecBCD-KO_5HA_long_rv and CAT_RecBCD-KO_5HA_long_fw) and the other pair shorter. (CAT_RecBCD-KO_5HA_short_rv and CAT_RecBCD-KO_5HA_short_fw).</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td> | <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td> | ||
| − | <td> | + | <td> We thought the band might be what we wanted but because of how few DNA the extraction yielded, these samples were not used in further experiments. |
| + | |||
| + | <figure class="figure"> | ||
| + | <img src="https://static.igem.org/mediawiki/2018/b/bd/T--munich--20180605_CAT-KO.jpg" alt=" "> | ||
| + | <figcaption class="figure-caption"></figcaption> | ||
| + | </figure> | ||
| + | |||
</td> | </td> | ||
</tr> | </tr> | ||
Latest revision as of 00:13, 18 October 2018
Transforming E. Coli DH5a to Amplify pKD3 for pRED/ET Engineering
2018/05/28| Participants: | Dominic Schwarz |
| Protocol: | Electro-transformation |
| Notes: | No notes. |
| Results: | No colonies, no growth; We decided to use a resistance cassette from pSB1C3 without FRT sites. |
Amplifying a Selection Cassette from pSB1C3
2018/05/29| Participants: | Enikö Baligács |
| Protocol: | PCR, Agarose gel, Gel extraction |
| Notes: | PCR with two primer pairs were tested to amplify a chloramphenicol resistance cassette for the knockout. Both pairs have a 50 bp overhang homology region to the knockout target in E.Coli genomes. One pair has longer annealing sequence (CAT_RecBCD-KO_5HA_long_rv and CAT_RecBCD-KO_5HA_long_fw) and the other pair shorter. (CAT_RecBCD-KO_5HA_short_rv and CAT_RecBCD-KO_5HA_short_fw). |
| Results: | We thought the band might be what we wanted but because of how few DNA the extraction yielded, these samples were not used in further experiments.
|