Note Book - iGEM Bordeaux 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.

As IB digestion didn't worked we did and tested it again

We then transformed the ligations into two new competent cells batches
One heat shock competent DH10B stock and one heat shock competent DH5α stock
We also tested the competence of the two new stocks with 1μg pBlueScript

Day 27 : 09-08-2012

Nothing on the plates ... Except for the DH5α pBluescript test