chimeric race products???

Gregor st2153 at zi.biologie.uni-muenchen.de
Thu Sep 23 05:16:01 EST 1999


Hi everybody!

I´ve tried to race several genes with the Marathon kit. I do nested PCRs
with little or no product in the first round (30 cycles) and many bands
and a smear in the second round 30 cycles as well (with recommended
temperatures 72 X5/70 X5/68 X20). the bands I´ve sequenced so far often
start with the expected bases but then the sequence "hops" to another -
totaly unrelated - sequence.

Do I have to lower (raise) temperatures?

Or lower/raise template concentration?

What are - generally - the parameters that favor for chimeric PCR
products/jumping PCR?


THanks for advice!!

Gregor
-- 
Gregor Bucher, LMU München
st2153 at zi.biologie.uni-muenchen.de

the problem with working on the cutting edge is 
that you get sliced from time to time



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