Difference between revisions of "Team:UCopenhagen/Notebook"

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<h4 class="media heading">Wet Lab Overview</h4>
 
<h4 class="media heading">Wet Lab Overview</h4>
 
<p>
 
<p>
−
Started making plates and initial transformations
+
Making plates, initial transformations, ordered more primers.
 
</p>
 
</p>
 
 
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<h4 class="media heading">Interdependency</h4>
 
<h4 class="media heading">Interdependency</h4>
 
<p>
 
<p>
−
Made glycerol stock of MG1655
+
Made glycerol stock of MG1655. Designed primers: yddG +/- his-tag, and pointmutations in trpE and aroG
 +
 
 
</p>
 
</p>
 
</div>
 
</div>
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<h4 class="media heading">Protein Import</h4>
 
<h4 class="media heading">Protein Import</h4>
 
<p>
 
<p>
−
Text
+
Inoculation and creation of glycerol stocks and streaking on LB plates of 8 bacterial cultures: Micrococcus luteus, Staphylococcus epidermis, Bacillus Cereus, Bacillus licheniformis, Serratia marcescens, Pseudomonas putida and Rhizobium meliloti.
 +
Preparation of additional LB plates without antibiotica.
 +
Creation of competent Mach1 cells and fast transformation with pRSET A to test efficiency. 
 +
 
 
</p>
 
</p>
 
</div>
 
</div>
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<h4 class="media heading">Interdependency</h4>
 
<h4 class="media heading">Interdependency</h4>
 
<p>
 
<p>
−
Made glycerol stock of MG1655
+
PCR from MG1655 stock. Amplified aroG #1 and #2 in first attempt, and trpE #2 at a higher annealing temp, then began optimizing the PCR for yddG and trpE#2 using gradient temperatures without success.
 
</p>
 
</p>
 
</div>
 
</div>
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<h4 class="media heading">Protein Import</h4>
 
<h4 class="media heading">Protein Import</h4>
 
<p>
 
<p>
−
Text
+
Fluorescent microscopy of S. marascens, E. coli and S. epidermis stained with bodipu.
 +
Removal of XbaI sites in pRSET A mod, PCR amplification and gel-extraction of the plasmid.
 +
Transformation of competent cells with pRSET amod 1, and test of XbaI site removal by digestion.
 +
Started work on pRSETjin and pRSETjin CPP vectors from the pRSET vector without XbaI site.
 +
 
 
</p>
 
</p>
 
</div>
 
</div>

Revision as of 08:59, 19 September 2017

N O T E B O O K

Week 1 (June 26 - July 2)

  • Wet Lab Overview

    Ordered primers, checked cell stocks and prepared LB plates



Week 2 (July 3 - July 9)

  • Wet Lab Overview

    Making plates, initial transformations, ordered more primers.

    Interdependency

    Made glycerol stock of MG1655. Designed primers: yddG +/- his-tag, and pointmutations in trpE and aroG

    Number Control

    Text

    Protein Import

    Inoculation and creation of glycerol stocks and streaking on LB plates of 8 bacterial cultures: Micrococcus luteus, Staphylococcus epidermis, Bacillus Cereus, Bacillus licheniformis, Serratia marcescens, Pseudomonas putida and Rhizobium meliloti. Preparation of additional LB plates without antibiotica. Creation of competent Mach1 cells and fast transformation with pRSET A to test efficiency.



Week 3 (July 10 - July 16)

  • Wet Lab Overview

    Started making plates and initial transformations

    Interdependency

    PCR from MG1655 stock. Amplified aroG #1 and #2 in first attempt, and trpE #2 at a higher annealing temp, then began optimizing the PCR for yddG and trpE#2 using gradient temperatures without success.

    Number Control

    Text

    Protein Import

    Fluorescent microscopy of S. marascens, E. coli and S. epidermis stained with bodipu. Removal of XbaI sites in pRSET A mod, PCR amplification and gel-extraction of the plasmid. Transformation of competent cells with pRSET amod 1, and test of XbaI site removal by digestion. Started work on pRSETjin and pRSETjin CPP vectors from the pRSET vector without XbaI site.