direct PCR from bacterial colonies.

Neil Percy 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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