PCR Frustrations

Dan Lasser dml1 at columbia.edu
Tue Feb 13 17:14:26 EST 1996


I am trying to optimize a hemi-nested DNA PCR protocol. My problem is 
smearing of lanes on the resulting gels with certain of the primers. 
Others give crisp bands. 

I have tried temperature otpimization, have cut back on dNTP and TAQ 
concentrations and have even added DMSO. Still smearing with occasional 
bands buried in the smear. The DNA sample amplifies well with other 
primers. Any suggestions?



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