Storage for PCR
ez005139 at chip.ucdavis.edu
Fri Aug 12 12:27:20 EST 1994
We are currently sampling zooplanktonic veliger larvae in the field,
formalin killing them and transfering them to denatured alcohol for PCR.
We are getting some success in amplification of a mtDNA region but would like
to improve this to 100%. We get 100 % success with purified mtDNA samples,
and pro-k digests of fresh single larvae - so we know the PCR is not the
source of failures. We are checking the possibility that the samples
are not our species, but the working hypothesis is that some degredation of
sample may be occuring.
Could anyone offer methods on how to easily prepare field samples of
marine planktonic larvae so that we can transport them back to the lab,
store them for a year or more, and then perform PCR? Does denatured alcohol
work as well as non-denatured alcohol (which is difficult to transport)?
Our formalin is dilute and quickly rinsed , but perhaps formalin is the
problem? Is there degredation with long term storage at -20 ?
Any advice will be greatly appreciated...
Bodega Marine Lab
P.O. Box 247
Bodega Bay, Ca.
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