help! PCR troubles..

gsbs1028 at UTSPH.SPH.UTH.TMC.EDU gsbs1028 at UTSPH.SPH.UTH.TMC.EDU
Sun Nov 7 19:57:14 EST 1993


I am (as yet unsuccessfully) to amplify across a 3.2 kb intron in
human genomic DNA. I am trying formamide and glycerol additions
(2.5% and 10%, respectively) and a hot start adding dNTPS at 70
deg. extension time is 3 min, also tried 3 min,.  with 5 sec
added on each successive cycle. this shouldn't be as tough
as it's going- can anyone out there give me some hints?
has anyone out there done a fragment this large or larger?
thanks!



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