Efficient method for analysis of recombinants?

Greg Nattrass gnattras at waite.adelaide.edu.au
Wed Feb 5 07:17:25 EST 1997


We use a technique called colony cracking to analyse recombinants.
Individual colonies are picked from a plate with a yellow tip, streaked
on a master plate and the remainder lysed in a solution containing
SDS/NaOH/EDTA. After 15 minutes @ 65C the lysate is run on a 10ml
minigel. A colony from the vector only plate is run as a size standard.
The procedure takes 40 minutes, recombinants are selected from the
master plate a few hours later and cultured. I have identified 200bp
inserts in pBlue using a 0.8% gel. For more information on the lysis
buffer etc, feel free to email me.

Greg Nattrass



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