Decontamination of polymerase chain reaction reagents using DEAE-cellulose.
Polymerase chain reaction (PCR) is widely used to detect specific DNA sequences for purposes of microbial identification, clinical diagnosis, and basic research. The most pernicious problem plaguing this technique is contamination of PCR reagents with previously amplified material. We propose a useful tool for PCR reagent purification from contaminating nucleic acid using DEAE-cellulose and present the analysis of this technique for both decontamination efficiency and an effect on the reagent activity. We also show the suitability of the proposed approach for decontamination of the Taq polymerase, monoclonal antibodies to Taq polymerase, and Moloney murine leukemia virus (M-MLV) reverse transcriptase.
Glushkov
S A
SA
Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences, Novosibirsk 630090, Russian Federation. sg23@mail.ru
Bragin
A G
AG
Dymshits
G M
GM
D003636:DEAE-Cellulose / Q000737:chemistry*; D003666:Decontamination; D007202:Indicators and Reagents / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D019914:Taq Polymerase / Q000378:metabolism
D007202:Indicators and Reagents; D003636:DEAE-Cellulose; D019914:Taq Polymerase