see DNA incubation time 11-14-12

2014-08-29

azim58 - see DNA incubation time 11-14-12


Does it make a difference to pipette the cells with DNA, leave the DNA
with the cells for 1 min, or use the cells immediately (1-10 s) after
adding DNA when doing an electroporation?

From my experience so far the answer is no.
On 11-8-12 I did an experiment to test this issue

10 ng DNA
1000 uL SOC
40 min 37 C incubation

1 min sample
7 colonies on 10^0 plate
  1. 4e4 cfu/ug

Pipetting the DNA
9 colonies on 10^0 plate
  1. 8e4 cfu/ug

10 s
about the same number of colonies as "1 min" and "pipetting", but it was
hard to count exactly since the agar was scratched.
about 1.5e4 cfu/ug