3' overlaps in PCR

DMEIER at mis.mcw.edu DMEIER at mis.mcw.edu
Mon Dec 14 18:04:06 EST 1992


Hello All PCR-ers,
  Two potential primers have a 3 base overlap at the 3' end (GGC), 
and have Tm's of 50.9 and 51.2 degrees C.  If I remove a G from, the 
forward primer for example, I get Tm's of 46.1 and 51.2 degrees C with 
a 2 base overlap at the 3' end (GG).  Is it worth sacrificing Tm 
compatibility for reducing 3' overlapping?  Incidently, the Tm of the 
target cDNA is 86.8 degrees C.
  Any comments will be appreciated.

  Thanks.

  Dan
  DMEIER at MIS.MCW.EDU



More information about the Methods mailing list