Elimination of bacterial DNA from Taq DNA polymerases by restriction endonuclease digestion.
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.
Carroll
N M
NM
Department of Clinical Ophthalmology, The Institute of Ophthalmology, London EC1V 9EL, United Kingdom.
Adamson
P
P
Okhravi
N
N
D004269:DNA, Bacterial / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D019914:Taq Polymerase / Q000032:analysis*
D004269:DNA, Bacterial; D019914:Taq Polymerase