Team:Evry/Notebook/w2

From 2013.igem.org

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<h2> Monday, June 24th </h2>
<h2> Monday, June 24th </h2>
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<p> Our competents cells made on friday the 21st were plated and incubated the whole week-end at 30°C. Today, we analyzed the plates and came to the conclusion that the medium was contaminated due to poor protocol execution. As a consequence, we decided to make new LB medium and recalculate the antibiotic concentrations.</p>
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<p> Our competents cells made on friday the 21st were plated and incubated the whole week-end at 30°C. Today, we analyzed the plates and came to the conclusion that the medium was contaminated due to poor protocol execution. As a consequence, we decided to make new LB medium and recalculate the antibiotic concentrations.<br/>
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New competent cells have been made for all three strains: DH5-alpha, BL21 and TOP10.</p>
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<b><div align="center">Protocol for LB medium preparation</b><br/></div>
<b><div align="center">Protocol for LB medium preparation</b><br/></div>
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Add 20 g of LB broth into 1000 mL of desalted water and autoclave.<br/>
Add 20 g of LB broth into 1000 mL of desalted water and autoclave.<br/>
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<b><div align="center">Protocol for LB medium preparation</b><br/></div>
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<b><div align="center">Protocol for antibiotic solutions preparation</b><br/></div>
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<p>New competents cells will be made.</p>
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<h2>Tuesday, June 25th</h2>
<h2>Tuesday, June 25th</h2>

Revision as of 16:16, 28 July 2013

Iron coli project

Week 2: 24th June - 30th June

Monday, June 24th

Our competents cells made on friday the 21st were plated and incubated the whole week-end at 30°C. Today, we analyzed the plates and came to the conclusion that the medium was contaminated due to poor protocol execution. As a consequence, we decided to make new LB medium and recalculate the antibiotic concentrations.
New competent cells have been made for all three strains: DH5-alpha, BL21 and TOP10.


Protocol for LB medium preparation

Add 20 g of LB broth into 1000 mL of desalted water and autoclave.


Protocol for antibiotic solutions preparation

Tuesday, June 25th

New competent cells have been made with the same strains of bacteria. We thought that the contamination of our cells was due either to the solution of CaCl2 that we used which may had not been sterile and/or incorrect manipulations. This time, we prepared new CaCl2 solutions that we made sure to autoclave correctly. Then, same tests as we did before of contamination and competence have been made.

Wednesday, June 26th

The DH5a strain was not contaminated and competent. The TOP10 strain was contaminated only on the Kanamycin petri dish and competent. The BL21 strain was contaminated and not usable.

Thurdsay, June 27th

We designed the plasmid constructions and primers we will use.