PCR problem:inverted repeats

Beth Wapelhorst weazel at seanet.com
Fri Jan 13 21:26:26 EST 1995


5gf4srivasta at vms.csd.mu.edu (NAMRATA SRIVASTAVA) writes:

I would maybe suggest adding 10% DMSO to the reaction.
(final concentration is 10%)

I know that it sounds like a lot, but this worked for me when I was 
trying to amplify a 1 kb product.

The DMSO keeps things denatured so that they can extend the whole way.

sugarbeth
weazel at seanet.com

>HI netters!
>	Im trying to PCR a 3-4 Kb DNA fragment which contains 1.6 Kb of 
>inverted repeat sequence. I get a shorter product which would correspond 
>to a secondary DNA structure loop (DNA hybridizing to itself) hence the taq
>is unable to read through the entire template. I have tried hot starts to
>eliminate this but to no avail. Does any one out there have any suggestions?
>				
>				Any helpful hint would be appreciated!
>						Thanx.
>						Namrata Srivastava
>						Marquette University.



More information about the Methods mailing list