Bacterial DNA removing question

Malay curiouser at ccmb.ap.nic.in
Mon Mar 8 03:04:04 EST 1999


>Can somebody help me with bacterial DNA removing from PCR mixes ?
>Taq DNA polymerase is often contaminated of EColi DNA. I've heard
>of method with 5-methoxypsoralene and near UV, but I don't know how
>to clear mine solution.

May I suggest some literatures in this regard:

Reliability of PCR decontamination systems.
PCR Methods Appl. 1994 Oct;4(2):117-23.

Eliminating PCR contamination: is UV irradiation the answer?
J Virol Methods. 1991 Aug;33(3):375-82.

Parameters affecting susceptibility of PCR contamination to UV inactivation.
Biotechniques. 1991 May;10(5):590-4.

Use of UV irradiation to reduce false positivity in polymerase chain
reaction.
Biotechniques. 1991 Apr;10(4):442, 444, 446.

Always have single primer and a negative control without the template DNA
for your PCR to check for contamination. I personally have never tried any
of the approach!  Most of the time it is easier to use a different stock of
enzyme or primers than trying to decontaminate! So, you are on your own!

Malay

Malay Kumar Basu
Centre for Cellular and Molecular Biology
Hyderabad 500007
I N D I A

Fax: (00-91)40-7171195
Phone: (00-91)40-7172241
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curiouser at ccmb.ap.nic.in







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