Optimal Conditions and Validation of Single-strand Conformation Polymorphism Technology for the Analysis of Microbial Communities in Activated Sludge
- VernacularTitle:SSCP技术分析活性污泥微生物群落结构的条件优化及检验
- Author:
Ai-Jie WANG
;
Hong-Jing KAN
;
Zhen-Guo YU
;
Nan-Qi REN
;
Chun-Shuang LIU
;
Yun-Qing ZHANG
;
Yang-Guo ZHAO
;
- Publication Type:Journal Article
- Keywords:
Single-strand conformation polymorphism, Optimal conditions, Validation, Microbial community
- From:
Microbiology
2008;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
Single-Strand Conformation Polymorphism(SSCP) is an effect method for investigating environment microbial genetic polymorphism, with its characterization of rapidness, simplicity, and sensitivity. However, many factors can influence the results of SSCP in the analysis of complex environment samples, and its optimization is highly needed. In this paper, optimal PCR-SSCP conditions were discussed based on PAGE concentration, formamide deionized in denaturing loading buffer, electrophoresis time and temperature. The resluts showed that the optimal conditions were as follows: 16S rDNA V1~V3 was selected as the targeted gene, the ratio of acrylamide to N, N-dimethylacrylamide in 12% polyacrylamide gel electrophoresis(PAGE)gel was 49:1, the ratio of formamide deionized in denaturing loading buffer was 1:3, running the SSCP gel at 300 V for 18 h (under 4 ℃). Aside from this, the validations using samples from a simultaneous desulfurification and denitrification bioreactor were conducted under this optimal conditions.
