Elimination of bacterial DNA from Taq DNA polymerases by restriction endonuclease digestion

被引:72
作者
Carroll, NM [1 ]
Adamson, P [1 ]
Okhravi, N [1 ]
机构
[1] Inst Ophthalmol, Dept Clin Ophthalmol, London EC1V 9EL, England
基金
英国惠康基金;
关键词
D O I
10.1128/JCM.37.10.3402-3404.1999
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The incidence of false positives due to the presence of bacterial DNA in Tag 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.
引用
收藏
页码:3402 / 3404
页数:3
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