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inhibitet PCR reaction

inhibitet PCR reaction - Protocols and Methods Forum

inhibitet PCR reaction - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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  #1  
Old 10-31-2007, 02:48 PM
Omar Hamarsheh
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Default inhibitet PCR reaction



Dear Colleges,

I am working with ITS1 PCR using Leishmania DNA samples extracted from
filter papers, the samples where originated from skin scraping and
contain blood. I used inhibitor control for each sample . few samples
were worked and gave positive. most of the others show no PCR bands,
the inhibition control sho also no band which mean that there is an
inhibition of the PCR reaction probably caused from something in
blood.
I would like to ask colleges who are working with clinical samples if
they have an inhibitted PCR reaction what could be done in this
regard.

thanks alot in advance for any help or suggestion.

Omar Hamarsheh

Charite school of Medicine
Dorotheenstr. 96
10117 Berlin
Germany

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  #2  
Old 11-01-2007, 03:17 PM
chovek69
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Default inhibitet PCR reaction

On Oct 31, 5:48 pm, "Omar Hamarsheh" <[Only registered users see links. ]> wrote:

Hi I have quite a bit experience with blood inhibited PCRs. The main
inhibitor is the hem from the hemoglobine but there are others. The
best remedy so far is the addition of nonacetylated Bovine Serum
Albumine (BSA) from Sigma, in 0.1 to 0.8 g/L final concentration.
Put in the PCR mix and sing. It depends also from the Taq but in
general all works the same. There are other polymerases that are not
inhibeted from the blood but they are expensive. (see for example this
[Only registered users see links. ]

Hope this helps
Regards


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