Colony PCR Failures?

Dom Spinella dspinella at chugaibio.com
Fri Dec 12 10:33:02 EST 1997


Hi All:

Lots of folks have been responding to my previous posting

http://www.bio.net/hypermail/METHDS-REAGNTS/9712/0311.html

in which I hypothesized that failures of colony PCR may be related to
host strain or plasmid vector and offered to tabulate and post the
results of replies from people who have had the technique succeed or
fail.  However, so far I am hearing only about the successes (heck --
maybe no one else has ever had the technique fail except me and Harry
Witchel!). But I suspect that some strains of bugs may be harder to
crack open (and may therefore require more harsh conditions to isolate
DNA than just throwing some bacterial debris directly into a PCR) or
perhaps some lower copy plasmids may be more difficult PCR templates. 
So how about it out there -- anyone else ever had problems with the
technique?  If so, what strains, plasmids, primers, and PCR conditions
have you tried?  Please cc my return e-mail if your reply -- I'd like to
post the final results early next week. Thanks to all who have already
replied. Is this a great forum or what!

-- Dom Spinella



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