Primer stocks that fail to PCR over time

Paul N Hengen pnh at fcsparc6.ncifcrf.gov
Wed Nov 23 16:02:01 EST 1994


Some netters have noticed a loss in the ability to amplify DNA by the
polymerase chain reaction when using the same lot of primers over a period of
time.  On occasion, the failure of PCR amplification was directly linked
to the primer stock going belly up for unknown reasons.

PCR primers initially worked shortly after being purchased, but after storage
for several months at -20 C as diluted stocks in water, these same primers
failed to give the same amount of amplified product in identical reactions.
Primers going bad also produced unreadable results in automated DNA sequencing
reactions using the dye terminator chemistry combined with cycle sequencing and
subsequent analysis on the ABI model 373A sequencer under the same conditions
that previously gave good results.  If new primers were purchased, the reactions
gave good results and acted as they did prior to storage at - 20 C.

I'm searching for people who have had the unfortunate experience of losing
their primer stocks to the big eppendorf tube in the sky when they've stored
them in their -20 C freezer. If you've had trouble with this problem and feel
like sharing your experience to be written in an upcoming TIBS column, please
post or e-mail. Thanks.

-Paul.

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* Paul N. Hengen, Ph.D.                           /--------------------------/*
* National Cancer Institute                       |Internet: pnh at ncifcrf.gov |*
* Laboratory of Mathematical Biology              |   Phone: (301) 846-5581  |*
* Frederick Cancer Research and Development Center|     FAX: (301) 846-5598  |*
* Frederick, Maryland 21702-1201 USA              /--------------------------/*
* - - -  Methods FAQ list -> http://www.ncifcrf.gov:2001/~pnh/info.html - - - *
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