title: An effective method for isolation of DNA from pig faeces and comparison of five different methods.
abstract: 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 beta-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.
authors.affiliation: Bioengineering Research Center for Animal Disease Prevention and Control, School of Life Science, Sichuan University, Chengdu, China.
authors.forename: Jun-ni
authors.initials: JN
authors.lastname: Tang
authors.forename: Zhi-guang
authors.initials: ZG
authors.lastname: Zeng
authors.forename: Hong-ning
authors.initials: HN
authors.lastname: Wang
authors.forename: Tai
authors.initials: T
authors.lastname: Yang
authors.forename: Peng-ju
authors.initials: PJ
authors.lastname: Zhang
authors.forename: Yu-ling
authors.initials: YL
authors.lastname: Li
authors.forename: An-yun
authors.initials: AY
authors.lastname: Zhang
authors.forename: Wen-qiao
authors.initials: WQ
authors.lastname: Fan
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhang
authors.forename: Xin
authors.initials: X
authors.lastname: Yang
authors.forename: Su-jun
authors.initials: SJ
authors.lastname: Zhao
authors.forename: Guo-bao
authors.initials: GB
authors.lastname: Tian
authors.forename: Li-kou
authors.initials: LK
authors.lastname: Zou
mesh_terms: D000818:Animals; D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D004275:DNA, Ribosomal / Q000235:genetics; D005243:Feces / Q000382:microbiology*; D005821:Genetic Techniques; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D013552:Swine
chemical_list: D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S