Advise on Inverse PCR
William_D_WARREN at UMAIL.UMD.EDU
Mon Mar 2 13:50:00 EST 1992
Is there anyone out there who has mastered the art of inverse PCR
(ie divergently oriented primers to amplify unknown flanking DNA) ?
I'm considering trying this and would like to know how difficult it
would be to amplify sequences of 2 or 3kb and perhaps as much as 5 kb?
Also, after recircularization, do you have to linearize the template DNA
or will closed circles work.
And another PCR question....I have just sequenced some PCR products that
were amplified with inosine containing primers and at each inosine I
have a G - I guess from this that Taq pol prefers to add a G when it
encounters inosine in the template. Is this valid ?????
Any info will be greatly appreciated :-)
CTR FOR AG BIOTECH
UNIVERSITY OF MARYLAND
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