Quntifying small amounts of DNA reliably
Thu Jan 30 16:17:28 EST 1997
mmd280 at sysc.abdn.ac.uk (s.pritchard) writes: > Hi
> I am working on LOH in lung cancer and have to use paraffin embedded tissue
> as the source material.The DNA is recovered from the tissue after standard
> Haemotoxylin/eosin staining .PCR is performed on this material with no further
> purification so as to minimise further degradation.The yields are obviously
> small and therefore I can not estimate the conc. by running it on a gel and
> i presume that such crudely prepared DNA would be useless for analysis on
> a spec. Any ideas?
> Thanks in advance
> Stuart Pritchard
> mmd280 at abdn.ac.uk
Stuart, I've used an Ethidium Bromide Assay where you spot a 1 ul aliquot of your
sample and also a 1 ul aliqout of a 1/10 dilution of your sample on a petri plate
that contains 0.8% agarose and EtBr. You also spot a set of standards of known
concentration (100, 75, 50, 25, 10 ng/ml). Let the spots dry for 10-15 minutes
and then photograph the plate under UV light. Then all you do is compare your
unknown to the standards. The agarose/EtBr plates can be prepared ahead of time
and stored in the fridge for one month. It may be worth a try. Mike.
Michael E. DeGraaf
Pharmacia and Upjohn Inc.
E-Mail: MEDEGRAA at AM.PNU.COM
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