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Rapid screening of recombinants


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4 replies to this topic

#1 praveen_iitb

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Posted 05 December 2004 - 08:42 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

#2 lcrab

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Posted 21 December 2004 - 08:37 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.

#3 DevGrp

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Posted 21 December 2004 - 08:42 AM

Yep I've always used colony PCR

#4 praveen_iitb

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Posted 22 December 2004 - 10:32 AM

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

#5 DevGrp

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Posted 23 December 2004 - 01:55 AM

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




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