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《Food Science》 2010-06
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Dilution-PCR and PCR-DGGE for Evolution of Effect of Pre-treatment Methods on Extraction of Bacterial DNA from Chilled Pork

JIANG Yun1,2,GAO Feng1,XU Xing-lian1,YE Ke-ping1,ZHOU Guang-hong1,*(1.National Center of Meat Quality and Safety Control,Nanjing Agricultural University,Nanjing 210095,China;2.Ginling College,Nanjing Normal University,Nanjing 210097,China)  
Dilution-PCR and PCR-DGGE were used to evaluate effect of four pre-treatment methods such as freeze-thaw,ultrasonification,shake and homogenization on extraction of bacterial DNA from chilled pork in this study.After 4 days of storage at 4 ℃,chilled pork was subjected to pre-treatment by the above methods for bacterial DNA extraction followed by dilution-PCR and PCR-DGGE analyses.Results showed that four pretreatment methods results in a slight difference in bacterial DAN yield but had no notable effect on PCR-DGGE analysis.Therefore,these pretreatment methods could be chosen according to actual requirement.Sequence analysis of DGGE bands of 16S rRNA gene V3 region showed that the predominant spoilage bacterium in chilled pork was Pseudomonas and others including Acinetobacter,Brochothrix,and Serratia were also detected.
【Fund】: 国家“863”计划重点项目(2008AA100804);; 江苏省高校自然科学基础研究项目(08KJB550005);; 上海市科委重大攻关项目(07dz19508)
【CateGory Index】: TS251.51
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