direct PCR from bacterial colonies.
saspercy at reading.ac.uk
Fri Aug 11 04:27:47 EST 1995
We routinely screen colonies by PCR as an alternative to minipreps. You
simply prepare a PCR supermix (50ng each primer, 0.5u Taq per reaction)
and pipette 20ul into each tube. Then using Gilson tip touch bacterial
colony and streak master plate, then drop tip into tube. When you have
picked desired number of colonies, remove the tips expelling all liquid
using finger. Cover with mineral oil and amplify. For upto 1kb
fragments I use 25 cycles (20sec 94, 20sec 55, 60sec 72). All done in
less than 2 hours! Hope this makes life easier.
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