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Journal of Clinical Microbiology, October 1999, p. 3402-3404, Vol. 37, No. 10
Department of Clinical Ophthalmology, The
Institute of Ophthalmology, London EC1V 9EL, United Kingdom
Received 2 April 1999/Returned for modification 4 June
1999/Accepted 6 July 1999
The incidence of false positives due to the presence of bacterial
DNA in Taq DNA polymerase is an obstacle to the use of PCR in the diagnosis of infection. We describe a method that uses a
restriction enzyme to destroy the ability of contaminating sequences to
act as templates for a nested PCR which uses primers based on the 16S
rRNA genes. The method was used prior to a PCR that amplified 10 fg of
bacterial DNA. This method can be readily adapted to suit other
sensitive PCRs required for clinical applications.
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Elimination of Bacterial DNA from Taq
DNA Polymerases by Restriction Endonuclease Digestion

*
Corresponding author. Mailing address: Department of
Clinical Ophthalmology, The Institute of Ophthalmology, Bath St.,
London EC1V 9EL, United Kingdom. Phone: 0171-608-6872. Fax:
0171-608-6931. E-mail:
nokhravi{at}menu.hgmp.mrc.ac.uk.
Present address: Department of Medical Biochemistry, University of
Stellenbosch, Tygerberg 7505, South Africa.
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