PCR sequencing problem

Walt Lilly C749SCB at SEMOVM.SEMO.EDU
Mon Mar 20 12:46:45 EST 1995


Hi,

We are trying to sequence a short (300bp) piece of DNA cloned by PCR.
We have isolated one band by toothpick PCR and have used this single
band as the template for sequencing.  The problem we are having is that
we are getting generally smeared lanes with some apparently redundant
bands showing in different lanes.  We also see the strongest bands
showing up at the very top of each sequencing lane.  Our control DNA
works perfectly, so this must be a problem with the template.
Any ideas?  Thanks in advance.


Lisa Gordon
Walt Lilly



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