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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