chimeric race products???

alex dobrovic adobrovic at medicine.adelaide.edu.au
Thu Sep 23 05:31:00 EST 1999


We have seen a lot of alternative splicing with RACE. Some of these
"unrelated sequences" are possibly introns.

.>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 M¸nchen
>st2153 at zi.biologie.uni-muenchen.de
>
>the problem with working on the cutting edge is
>that you get sliced from time to time


Alexander Dobrovic, Ph.D.
Chief Medical Scientist
Department of Haematology-Oncology
The Queen Elizabeth Hospital
Woodville, SA 5011, Australia

Tel 011 (or your countries' international code) 61-8-8222 7676
Fax 011 (or your countries' international code) 61-8-8222 6046
adobrovic at medicine.adelaide.edu.au





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