IGEM:Team Bordeaux/2009/Notebook/Bacterial eyespot/2012/07/10

Day 6

Every tube grown well overnight except for the two 1.B tubes wich are clear after centrifugation we confirmed that nothing grew on the two 1.B tubes

We did a miniprep for the other tubes , As usual we used a QIAprep® Spin Miniprep kit from QIAGEN® [1]

We checked the samples concentration with the nanodrop [2]

We still got bad results ( around 15ng/μL) except for one 1.D tube ( 38,3ng/μL) and one 5.B tube ( 28ng/μL)

We decided to run an agarose 1% gel electrophoresis to confirm what we got on the samples

1.A = BBa_I14015 + pSB2K3 = 4.6 kb

1.D = BBa_C0051 + pSB1A2 = 2.8 kb

5.B = BBa_B0015 + pSB1AK3 = 3.3 kb

We can see that the samples contain the good plasmids

The strange thing is that the concentration seems higher in the 1.A and 5.B samples

We got one sample of XL1blue strain competent cells to test

So we did an electroporation as usual[3] and took 1.B DNA to transform

We plated 50μL and 100μL the transformants on two LB + Ampicilin 100μg/mL plates

We then put them to incubate at 37°C overnight