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Team:Bordeaux/Sixth
From 2012.igem.org
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| - | <h4>Day 25 : | + | <h4>Day 25 : 07-08-2012</h4> |
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| - | <h4>Day | + | <h4>Day 26 : 08-08-2012</h4> |
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| - | Today we digested and | + | Today we did a miniprep with the IAIB precultures from yesterday We got poor results from 14ng/μL to 36,6ng/μL DNA on the tubes We runned simple digested samples on an agarose gel but we couldn't discriminate the difference between the vector only and the IAIB+vector samples due to a too small deferences We'd have to run a more concentrated gel. |
</br> | </br> | ||
| - | We then | + | We then tried the assembly of IAIB ICIE IIAIIB IIC(1)5B We digested the bricks and tested them on agarose gel. |
| + | <div class="img-box"> | ||
| + | <figure><a><img src="http://openwetware.org/images/a/a3/Restriction0808_Bricks.jpg" alt=""></a></figure> | ||
| + | </div> | ||
| + | </br> | ||
| + | As IB digestion didn't worked we did and tested it again | ||
| + | <div class="img-box"> | ||
| + | <figure><a><img src="http://openwetware.org/images/1/1a/Restriction0808_1B.png" alt=""></a></figure> | ||
| + | </div> | ||
| + | </br> | ||
| + | We then transformed the ligations into two new competent cells batches | ||
| + | </br> | ||
| + | One heat shock competent DH10B stock and one heat shock competent DH5α stock | ||
| + | </br> | ||
| + | We also tested the competence of the two new stocks with 1μg pBlueScript | ||
</p> | </p> | ||
Revision as of 06:45, 24 September 2012
Day 24 : 06-08-2012
As the transformation didn't gave any colonies on the plate We decided to try again , and control on agarose gel as the gel seemed promising we transformed 2 μL of plasmid DNA We also tester the cells competence by transforming 1μg of pBlueScript and pSB1A3.
Day 25 : 07-08-2012
No colonies on the plates again ... even on the two tests we clearly have a competence issue. So we need to make a new stock today we tested the biobricks from day 19 on agarose gels
Day 26 : 08-08-2012
Today we did a miniprep with the IAIB precultures from yesterday We got poor results from 14ng/μL to 36,6ng/μL DNA on the tubes We runned simple digested samples on an agarose gel but we couldn't discriminate the difference between the vector only and the IAIB+vector samples due to a too small deferences We'd have to run a more concentrated gel. We then tried the assembly of IAIB ICIE IIAIIB IIC(1)5B We digested the bricks and tested them on agarose gel.
