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Rapid screening of recombinants - (Dec/05/2004 )

HI,
any body knows a good method for rapid screening of recombinants to identify clone.
as most of the methods are time consuming and very expensive.
Thanx in advance
praveen

-praveen_iitb-

QUOTE (praveen_iitb @ Dec 5 2004, 09:42 AM)
HI,
any body knows a good method for rapid screening of recombinants to identify clone.
as most of the methods are time consuming and very expensive.
Thanx in advance
praveen

Try to use PCR.

-lcrab-

Yep I've always used colony PCR

-DevGrp-

HI Icerab & devgrp,
Thanx for ur reply , but dont u think this method is very expensive. Like if u have to screen 50 colonies or more. In this case what u will prefer.
bye

-praveen_iitb-

I would do 50+ 10ul PCR reactions, for us thats cheaper than the technicians / my time to do 50+ mini-preps (and the restriction enzymes for digests can be expensive) and if you do the colony PCR in the morning you can have positive colonies by the afternoon and even save a day.

The other alternative to screen "lots" of colonies is to do plate lifts onto membranes and then do hybridization but that can get expensive (32P / non radioactive detection)
good luck

-DevGrp-