Difference between revisions of "Team:Munich/engineering2.html"
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| − | <h4>Transforming E. Coli Rosetta and MG1655 for pRED/ET | + | <h4>Transforming E. Coli Rosetta and MG1655 for pRED/ET Engineering</h4> |
<em>2018/08/28</em> | <em>2018/08/28</em> | ||
<table class="table table-borderless"> | <table class="table table-borderless"> | ||
| Line 22: | Line 22: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td>Results:</td> | + | <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td> |
| − | <td> | + | <td>Colonies only on E. Coli MG1655 plate; |
No colonies on E. Coli Rosetta | No colonies on E. Coli Rosetta | ||
</td> | </td> | ||
| Line 29: | Line 29: | ||
</table> | </table> | ||
| − | <h4>pRED/ET | + | <h4>pRED/ET Genome Engineering of Delta msb-B and Delta recBCD Strains</h4> |
<em>2018/08/29</em> | <em>2018/08/29</em> | ||
<table class="table table-borderless"> | <table class="table table-borderless"> | ||
| Line 39: | Line 39: | ||
<tr> | <tr> | ||
<td>Protocol:</td> | <td>Protocol:</td> | ||
| − | <td>pRED/ET engineering | + | <td> |
| + | <a href="https://static.igem.org/mediawiki/2018/b/bf/T--Munich--pRED_ET-Ecoli_Gene_Deletion_Kit.pdf.pdf" target="_blank">pRED/ET engineering</a> | ||
| + | |||
| + | </td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Notes:</td> | <td>Notes:</td> | ||
| − | <td> | + | <td>The template for the resistance cassette for deletions was taken from a plasmid containing mRFP |
</td> | </td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td>Results:</td> | + | <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td> |
| − | <td> | + | <td>Red colonies. Elena (Advisor) and Dominic decided to repeat the experiment but to do a dpnI digest before to get rid of the template DNA. |
</td> | </td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
| − | <h4>Transforming E. Coli DH5a to | + | <h4>Transforming E. Coli DH5a to Find a Reason for the Contamination</h4> |
<em>2018/08/30</em> | <em>2018/08/30</em> | ||
<table class="table table-borderless"> | <table class="table table-borderless"> | ||
| Line 73: | Line 76: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td>Results:</td> | + | <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td> |
| − | <td> | + | <td>Red colonies on all plates -> mRFP contamination |
</td> | </td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
| − | <h4>Creating a | + | <h4>Creating a Selection Cassette from pSB1C3</h4> |
<em>2018/08/30</em> | <em>2018/08/30</em> | ||
<table class="table table-borderless"> | <table class="table table-borderless"> | ||
| Line 100: | Line 103: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td>Results:</td> | + | <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td> |
<td>CAP_recBCD 18ng/ul | <td>CAP_recBCD 18ng/ul | ||
CAP_msbB 12ng/ul | CAP_msbB 12ng/ul | ||
| Line 107: | Line 110: | ||
</table> | </table> | ||
| − | <h4>pRED/ET | + | <h4>pRED/ET Genome Engineering of Delta msb-B and Delta recBCD Strains</h4> |
<em>2018/09/01</em> | <em>2018/09/01</em> | ||
<table class="table table-borderless"> | <table class="table table-borderless"> | ||
| Line 117: | Line 120: | ||
<tr> | <tr> | ||
<td>Protocol:</td> | <td>Protocol:</td> | ||
| − | <td>pRED/ET engineering | + | <td><a href="https://static.igem.org/mediawiki/2018/b/bf/T--Munich--pRED_ET-Ecoli_Gene_Deletion_Kit.pdf.pdf" target="_blank">pRED/ET engineering</a></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Notes:</td> | <td>Notes:</td> | ||
| − | <td> | + | <td>The template for the resistance cassette for deletions was taken from a plasmid containing mRFP |
</td> | </td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td>Results:</td> | + | <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td> |
| − | <td> | + | <td>Colonies on both plates |
</td> | </td> | ||
| Line 132: | Line 135: | ||
</table> | </table> | ||
| − | <h4>Verifying | + | <h4>Verifying Deletion Strains of E. Coli MG1655</h4> |
<em>2018/09/05</em> | <em>2018/09/05</em> | ||
<table class="table table-borderless"> | <table class="table table-borderless"> | ||
| Line 159: | Line 162: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td>Results:</td> | + | <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td> |
| − | <td> | + | <td>All 5 picked colonies were positive for the insertion of the selection cassette PIC |
</td> | </td> | ||
</tr> | </tr> | ||
Latest revision as of 00:25, 18 October 2018
Transforming E. Coli Rosetta and MG1655 for pRED/ET Engineering
2018/08/28| Participants: | Dominic Schwarz |
| Protocol: | Electrocompetent transformation |
| Notes: | Genetic engineering is planned in E. Coli Rosetta, MG1655 is E.Coli WT as backup. |
| Results: | Colonies only on E. Coli MG1655 plate; No colonies on E. Coli Rosetta |
pRED/ET Genome Engineering of Delta msb-B and Delta recBCD Strains
2018/08/29| Participants: | Dominic Schwarz |
| Protocol: | pRED/ET engineering |
| Notes: | The template for the resistance cassette for deletions was taken from a plasmid containing mRFP |
| Results: | Red colonies. Elena (Advisor) and Dominic decided to repeat the experiment but to do a dpnI digest before to get rid of the template DNA. |
Transforming E. Coli DH5a to Find a Reason for the Contamination
2018/08/30| Participants: | Dominic Schwarz |
| Protocol: | Electrocompetent transformation |
| Notes: | CAP_recBCD, CAP_msbb, psb1c3_mrfp in Dh5a |
| Results: | Red colonies on all plates -> mRFP contamination |
Creating a Selection Cassette from pSB1C3
2018/08/30| Participants: | Dominic Schwarz |
| Protocol: | Restrition digest, PCR purification |
| Notes: | DpnI |
| Results: | CAP_recBCD 18ng/ul CAP_msbB 12ng/ul |
pRED/ET Genome Engineering of Delta msb-B and Delta recBCD Strains
2018/09/01| Participants: | Dominic Schwarz |
| Protocol: | pRED/ET engineering |
| Notes: | The template for the resistance cassette for deletions was taken from a plasmid containing mRFP |
| Results: | Colonies on both plates |
Verifying Deletion Strains of E. Coli MG1655
2018/09/05| Participants: | Dominic Schwarz |
| Protocol: | Colony PCR, Agarose gel |
| Notes: | Primers: VF2, geno_msb-B_rv, geno_recBCD_rv;
Ta: 48°C
t= 1kb/min RecBCD: 1,2,3,4,5 msbB: 35,36,37,38,39 expected: RecBCD 443bp Expected: msb-B 574bp |
| Results: | All 5 picked colonies were positive for the insertion of the selection cassette PIC |
