AFLP troubleshooting

Tim Jackson tim.jackson at nrc.ca
Wed May 16 11:37:11 EST 2001


i'm using the GibcoBRL AFLP kit on Atlantic halibut.  i'm visualising my
product on an ABI 377 (fluorescent labelled EcoRI primers).  mostly
things are working very well except that my signal intesity for bands >
350 bp drops way off (i assume because of decreased amt. of PCR product
for the larger fragments).  i tried a few different Taq's and settled on
one which helped improve my signal from 200 to 350 bp but haven't been
able to make any improvements beyond that.  i thought i might try
increasing my extension time (currently 94 for 30s, 56 for 30s and 72
for 60s).
anyone have any bright ideas???

thanks!

tim




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