Elimination of Bacterial DNA from Taq DNA Polymerases by Restriction Endonuclease Digestion

AUTOR(ES)

Carroll, Nora M.

FONTE

American Society for Microbiology

RESUMO

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.

Documentos Relacionados

• Differentiation of Acanthamoeba strains from infected corneas and the environment by using restriction endonuclease digestion of whole-cell DNA.
• Identification of Campylobacter coli isolates from animals and humans by bacterial restriction endonuclease DNA analysis.
• Oligonucleotide-directed restriction endonuclease digestion.
• Cytosine methylated DNA synthesized by Taq polymerase used to assay methylation sensitivity of restriction endonuclease HinfI.
• Comparison of the Restriction Endonuclease Digestion Patterns of Mitochondrial DNA from Normal and Male Sterile Cytoplasms of ZEA MAYS L