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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2021.12
- 수록면
- 783 - 790 (8page)
- DOI
- 10.1007/s11816-021-00723-z
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초록· 키워드
오류제보하기
Polymerase chain reaction (PCR) is one of the most commonly used methods in diagnostic and molecular biology. However, DNA contamination can lead to incorrect results. This poses a major problem, especially in the monitoring of genetically modified organisms (GMOs). We investigated DNA contamination in 16 commercial Taq DNA polymerase reagents. Many of the Taq DNA polymerase reagents were contaminated with the 16S rRNA gene and an ampicillin resistance gene (bla). We attempted to decontaminate the Taq DNA polymerase reagents using UV, γ-ray, and electron-ray irradiation, as well as nylon membrane and FTA card treatment. UV irradiation reduced the activity of Taq DNA polymerase. Various wavelengths of γ-ray and electron-ray irradiation had no decontamination effect. However, 24-h treatment with three nylon membrane disks removed DNA contamination. FTA card treatment, which has membrane-like functions, also removed decontamina- tion but decreased enzyme activity. We further confirmed that the nylon membrane-treated Taq DNA polymerase reagent could be used in the detection of the bla and NPTII genes in genetically modified E. coli. These results are useful for future monitoring of GMOs.
목차
등록된 정보가 없습니다.
참고문헌 (25)
참고문헌 신청
Beckers T / 1994 / Highly sensitive northern hybridization of rare mRNA using a positively charged nylon membrane / Biotechniques 16(6):1074~1078
Böttger EC / 1990 / Frequent contamination of Taq polymerase with DNA / Clin Chem 36(6):1258~1259
Carroll NM / 1999 / Elimination of bacterial DNA from Taq DNA polymerases by restriction endonuclease digestion / J Clin Microbiol 37(10):3402~3404
Champlot S / 2010 / An efficient multistrategy DNA decontamination procedure of PCR reagents for hypersensitive PCR applications / PLoS ONE 5(9):e13042
Chiang CS / 2005 / Presence of beta-lactamase gene TEM-1 DNA sequence in commercial Taq DNA polymerase / J Clin Microbiol 43(1):530~531
Böttger EC / 1990 / Frequent contamination of Taq polymerase with DNA / Clin Chem 36(6):1258~1259
Carroll NM / 1999 / Elimination of bacterial DNA from Taq DNA polymerases by restriction endonuclease digestion / J Clin Microbiol 37(10):3402~3404
Champlot S / 2010 / An efficient multistrategy DNA decontamination procedure of PCR reagents for hypersensitive PCR applications / PLoS ONE 5(9):e13042
Chiang CS / 2005 / Presence of beta-lactamase gene TEM-1 DNA sequence in commercial Taq DNA polymerase / J Clin Microbiol 43(1):530~531
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