Advise on Inverse PCR

ww40 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 :-)

BILL WARREN
CTR FOR AG BIOTECH
UNIVERSITY OF MARYLAND




More information about the Methods mailing list