- Decontamination of polymerase chain reaction reagents using DEAE-cellulose.
Decontamination of polymerase chain reaction reagents using DEAE-cellulose.
Analytical biochemistry (2009-06-16)
S A Glushkov, A G Bragin, G M Dymshits
PMID19523915
摘要
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.
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二乙氨基乙基-葡聚糖®, aqueous ethanol suspension, 40-160 μm (wet), exclusion limit ~1,000,000 Da