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王红宁

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期刊论文

An effective method for isolation of DNA from pig faeces and comparison of five different methods

王红宁Jun-ni Tang Zhi-guang Zeng Hong-ning Wang * Tai Yang Peng-ju Zhang Yu-ling Li An-yun Zhang Wen-qiao Fan Yi Zhang Xin Yang Su-jun Zhao Guo-bao Tian Li-kou Zou

Journal of Microbiological Methods 75(2008)432-436,-0001,():

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摘要/描述

Polymerase chain reaction (PCR) detection of microorganism in faecal specimens is hampered by poor recovery of DNA and by the presence of PCR inhibitors. In this paper, we describe a new modified method for extracting PCR-quality microbial community DNA from pig faecal samples, which combines the pretreatment with polyformaldehyde, and subsequent DNA lysis in the presence of CTAB, salt, PVP, and β-mercaptoethanol, followed by isolation of nucleic acids using chloroform (no phenol) based protocol. The method resulted in a 1.3- to 11-fold increase in DNA yield when compared to four other widely used methods. Genomic DNA extracted from all five methods was assessed by both agarose gel electrophoresis and polymerase chain reaction for amplification of 16S rDNA specific fragments. The results showed that the improved method represented a reproducible, simple, and rapid technique for routine DNA extraction from faecal specimens and was notably better than using the QIAamp® DNA Stool Mini Kit.

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