Difference between revisions of "Team:Munich/engineering2.html"

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

@@ Line 2: / Line 2: @@
 <div class="event-info">
-<h4>Transforming E. Coli Rosetta and MG1655 for pRED/ET engineering</h4>
+<h4>Transforming E. Coli Rosetta and MG1655 for pRED/ET Engineering</h4>
 <em>2018/08/28</em>
 <table class="table table-borderless">
@@ Line 23: / Line 23: @@
   <tr>
        <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td>
-       <td>colonies only on E. Coli MG1655 plate;
+       <td>Colonies only on E. Coli MG1655 plate;
 		No colonies on E. Coli Rosetta
 </td>
@@ Line 29: / Line 29: @@
 </table>
-<h4>pRED/ET genome engineering of delta msb-B and delta recBCD strains</h4>
+<h4>pRED/ET Genome Engineering of Delta msb-B and Delta recBCD Strains</h4>
 <em>2018/08/29</em>
 <table class="table table-borderless">
@@ Line 39: / Line 39: @@
      <tr>
        <td>Protocol:</td>
-       <td>pRED/ET engineering protocol</td>
+       <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>
        <td>Notes:</td>
-       <td>the template for the resistance cassette for deletions was taken from a plasmid containing mRFP
+       <td>The template for the resistance cassette for deletions was taken from a plasmid containing mRFP
 </td>
      </tr>
   <tr>
        <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></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>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>
      </tr>
 </table>
-<h4>Transforming E. Coli DH5a to find a reason for the contamination</h4>
+<h4>Transforming E. Coli DH5a to Find a Reason for the Contamination</h4>
 <em>2018/08/30</em>
 <table class="table table-borderless">
@@ Line 74: / Line 77: @@
   <tr>
        <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td>
-       <td>red colonies on all plates -> mRFP contamination
+       <td>Red colonies on all plates -> mRFP contamination
 </td>
      </tr>
 </table>
-<h4>Creating a selection cassette from pSB1C3</h4>
+<h4>Creating a Selection Cassette from pSB1C3</h4>
 <em>2018/08/30</em>
 <table class="table table-borderless">
@@ Line 107: / Line 110: @@
 </table>
-<h4>pRED/ET genome engineering of delta msb-B and delta recBCD strains</h4>
+<h4>pRED/ET Genome Engineering of Delta msb-B and Delta recBCD Strains</h4>
 <em>2018/09/01</em>
 <table class="table table-borderless">
@@ Line 117: / Line 120: @@
      <tr>
        <td>Protocol:</td>
-       <td>pRED/ET engineering protocol</td>
+       <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>
        <td>Notes:</td>
-       <td>the template for the resistance cassette for deletions was taken from a plasmid containing mRFP
+       <td>The template for the resistance cassette for deletions was taken from a plasmid containing mRFP
 </td>
      </tr>
   <tr>
        <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td>
-       <td>colonies on both plates
+       <td>Colonies on both plates
 </td>
@@ Line 132: / Line 135: @@
 </table>
-<h4>Verifying deletion strains of E. Coli MG1655</h4>
+<h4>Verifying Deletion Strains of E. Coli MG1655</h4>
 <em>2018/09/05</em>
 <table class="table table-borderless">
@@ Line 160: / Line 163: @@
   <tr>
        <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td>
-       <td>all 5 picked colonies were positive for the insertion of the selection cassette PIC
+       <td>All 5 picked colonies were positive for the insertion of the selection cassette PIC
 </td>
      </tr>