extraction and PCR problem
clarosa at biocomp.unl.edu
Wed Aug 2 18:04:16 EST 2000
Valérie Moreau wrote:
> I extract DNA from whole mussels (0.2 - 0.7 mm) that have been in ethanol
> since October 1999. The extraction protocol is the usual phenol,
> phenol-chloroform-isoamylalcohol, and then chloroform-isoamylalcohol before
> ethanol precipitation.
> The problem is when I run PCR (target gene: glu-5 (242-380 bp)) with fresh
> or frozen mussel, everything works well. But with DNA extracts from mussels
> in ethanol, PCR doesn't work. It smears with no band.
> Could it be possible that ethanol preservation has a destructing effect on
> DNA structure? Is there a modified protocol for extracting DNA from ethanol
> preserved samples? Do I need to do something with the extracted DNA before
> runing PCR?
> Thank you very much.
> Valerie Moreau
> valerie.moreau at agr.gouv.qc.ca
I will consult my former boss who does preserved fish, upstairs....
Perhaps the ethanol does not really penetrate the tissues and inactivate
DNA degradation.??What is your ethanol preservation procedure?
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