Reasons for PCR failure....?

Riekeltje Koedood rkoedood at bio.bu.edu
Thu Nov 14 12:50:01 EST 1996


Another idea - My invitro transcription reactions often stopped
working after other people had used my reagents. Found out the 
reason - they thaw the NTP's at RT or sometimes even at 37 degrees.

As long as I always take care that my NTP stocks don't get too warm
(I thaw on ice, or quick thas in my hand and immediately put on ice),
my reactions keep working.

Marieke

David Demezas (demed at vms.ucc.okstate.edu.) wrote:
: We had some problems for a long time until one of my graduate students
: started looking at the storeage buffer of Taq pol.  There is a big
: difference in the storeage buffers.  Some storeage buffers have Triton
: X-100 and others have nonidet-P40 and Tween 20.  Our best results have
: been with those buffers with nonidet and tween which by the way I
: believe PE Taq is stored in this type of buffer.  Check your buffers;
: like you we tried everything-so we thought.



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