农业环境科学学报2011，Vol.30Issue(3)：599-604,6.
PCR-DGGE技术用于猪场沼气池细菌群落分析的条件优化
Optimization of Denaturing Gradient Gel Electrophoresis for Bacterial Communities in Biogas Slurry of Pig Farm
摘要
Abstract
Ploymersse chain reaction and denaturing gredient gel electrophoresis( PCR-DGGE ) technique was used to reveal the bacterial community composition and diversity associated with biogas slurry of pig farm. In order to improve the accuracy, sensitivity and repeatability of DGGE based on 16S rDNA V3 region, the running parameters of DGGE were optimized by level electrophoresis, electrophoresis time and dying time, respectively. The results showed that the best range of denaturing gradients was 35％～60％ in 8％ polyacrylamide gels, running time was 12 hours, and the dying time was 30 rain using SYBR Green Fluorescent Dye. Furthermore, bacterial community from 5 bingas digesters of pig farms was also performed by using these optimized reaction conditions. It was found that the 5 samples exhibited effective separation and rich microbial diversity. This indicated that different samples would result in the distinct change of microbial diversity and microbial community structure of biogas slurry. PCR-DGGE was proven to be a powerful tool for describing the bacterial flora in bingas slurry of pig farms. The results provide a basis for studying the bacterial community in livestock manure treatment.关键词
PCR-DGGE/猪场/沼气池/细菌群落Key words
PCR-DGGE/ pig farm/ biogas slurry/ bacteria communities分类
资源环境引用本文复制引用
魏勇,王红宁,廖党金,伍志伟,管仲斌..PCR-DGGE技术用于猪场沼气池细菌群落分析的条件优化[J].农业环境科学学报,2011,30(3):599-604,6.基金项目
四川省畜牧科学研究院基本科研业务费专项(SASA2009A06) （SASA2009A06）
四川生猪产业技术体系创新团队项目(sccxtd-001) （sccxtd-001）
省财政良种繁育专项(SASA2009YZ001) （SASA2009YZ001）
