PCR variability with DNA extraction method

Dr Claudia Macaubas macaubas at ichr.uwa.edu.au
Thu Aug 8 04:57:24 EST 1996


In article <3206084C.1652 at shef.ac.uk>, P R Orange <P.Orange at shef.ac.uk> wrote:

> Hello, I was wondering if any one else has experienced this problem and 
> if so, what they did to solve it.
> 
> I am using a PCR-restriction digestion assay to genotype some blood 
> samples, and I find that one of the polymorphisms is completely 
> undetectable when I use the "Nucleon" kit from Scotlab to extract the 
> DNA, but when I take the same blood sample, boil for 15 mins, spin out 
> the crud and use the supernatant, there is no problem in detecting this 
> polymorphism.
> 
> I've done extensive experiments to ensure that it is not a variability 
> in the PCR or digestion steps, and IT IS NOT,
> 
> Please can anyone help me??
> 
> Thanks in advance,
> 
> Paul Orange (P.Orange at sheffield.ac.uk)

Hi:

I had a similar problem when I was doing microsatellite typing. Samples
extracted with kits did not work so well as samples extracted with phenol.
Even when I tested samples sent to me, some extracted with a different kit
then mine, and some with phenol, the results were the same.

-- 
Claudia

TVW Telethon Institute for Child Health Research
(Company Limited by Guarantee)
Western Australia

Fax 61 9 388 3414



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