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

• Published in: Journal of microbiological methods Vol. 75; no. 3; pp. 432 - 436
• Main Authors: Tang, Jun-ni, Zeng, Zhi-guang, Wang, Hong-ning, Yang, Tai, Zhang, Peng-ju, Li, Yu-ling, Zhang, An-yun, Fan, Wen-qiao, Zhang, Yi, Yang, Xin, Zhao, Su-jun, Tian, Guo-bao, Zou, Li-kou
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01.12.2008; Elsevier
• Subjects: Animals; Bacteria - genetics; Bacteria - isolation & purification; Bacteriological methods and techniques used in bacteriology; Bacteriology; Biological and medical sciences; Comparison; DNA extraction; DNA, Bacterial - genetics; DNA, Bacterial - isolation & purification; DNA, Ribosomal - genetics; Feces - microbiology; Fundamental and applied biological sciences. Psychology; Genetic Techniques; Microbiology; PCR; Pig faeces; Polymerase Chain Reaction; RNA, Ribosomal, 16S - genetics; Swine; β-mercaptoethanol; Pig faeces; DNA extraction; β-mercaptoethanol; Comparison; PCR; Statistical analysis; Method; Pig; Polymerase chain reaction; Vertebrata; Mammalia; DNA; Bacteria; Artiodactyla; Feces; Ungulata
• ISSN: 0167-7012; 1872-8359
• DOI: 10.1016/j.mimet.2008.07.014

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.