Note Book - iGEM Bordeaux 2012

Day 10 : 16-07-2012

This week we decided to make our own competent cells
Regarding the bad/average results we had with our commercial DH5α and XL1blue we decided to try with DH10B strain
We followed the protocol proposed by the Read Protocol

Day 11 : 17-07-2012

Day 2 of making new competent cells Read Protocol
We stopped incubating our cells at an OD600nm of 0.31
In the end we made 192 tubes of 50μL competent cells.

Day 12 : 18-07-2012

The competence test gave good results with a good amount of colonies on each plates ( ≈ 300 ) and no colonies on the control plate.
Today we will transform DNA for every biobrick left.
We will use our new competent cells.

We will proceed by heat-shock as described in the protocol Read Protocol

Day 13 : 19-07-2012

Today we picked single colonies and put the to grow in two 5 mL LB + Ampicilin 100μg/mL
Except for 3.B who gave no colonies
We put the tubes to incubate overnight at 37°C

Day 14 : 20-07-2012

After 13 hours incubating we did a miniprep with the tubes. We got really poor results on the nanodrop for every biobrick (>3ng/μL). As we had really small cells pellets after the first centrifugation we suspect we didn't have enough cells in the beginning.