期刊文献+

Chelex-100法和碱性裂解法提取细菌DNA的比较 被引量:21

Comparison of methods for extraction of DNA by chelex-100 and alkali split
下载PDF
导出
摘要 目的:通过对两种DNA提取方法的比较,寻找一种灵敏、快速、经济实用的提取方法。方法:采用Chelex-100法和碱性裂解法分别提取不同浓度大肠杆菌ATCC25922和金黄色葡萄球菌ATCC25923的DNA,利用通用引物扩增16SrDNA片段检测两种方法的灵敏度。结果:碱性裂解法提取的不同浓度的两种细菌DNA,经PCR扩增均未显出目的条带;Chelex-100法提取不同浓度的两种细菌DNA均扩增出目的条带,PCR检测大肠埃希菌和金黄色葡萄球菌的灵敏度均为1.2×101cfu/ml,电泳结果提示相同浓度的大肠杆菌被提取到的DNA含量比金黄色葡萄球菌的含量高。结论:Chelex-100法操作简便,可用于细菌DNA的快速提取。 Objective: To find a sensitive,rapid and economic method of DNA extraction by comparison of Chelex-100 with alkali split.Methods:DNA from different concentrations of E.coli ATCC25922 and Staphylococcus aureus ATCC25923 was extracted using Chelex-100 and Alkali split methods.The extracted DNA was amplified using universal primers for 16SrDNA fragment.Results:No signal was obtained using PCR on the DNA samples extracted from different concentrations of two bacteria by alkali split.All DNA samples extracted from different concentrations of two bacteria by Chelex-100 were amplified.The sensitivity of PCR was 1.2×101 cfu/ml for either the detection of E.coli or Staphylococcus.The result of electrophoresis showed that the DNA levels were higher in E.coli than in Staphylococcus using the same concentration of bacteria for DNA extraction.Conclusion:The Chelex-100 extraction method is a simple and valuable method for rapid isolation of DNA from bacteria.
出处 《中国卫生检验杂志》 CAS 2008年第8期1565-1566,共2页 Chinese Journal of Health Laboratory Technology
关键词 Chelex-100 PCR 16SrDNA 碱性裂解法 Chelex-100 PCR 16SrDNA Alkali split
  • 相关文献

参考文献3

二级参考文献13

  • 1Li Y,Caufield PW. The fidelity of initial acquisition of mutants streptococci by infants from their mothers[J]. J Dent Res, 1995,74(2):681~685.
  • 2Li Y,Caufield PW. Arbitrarily primed polymerase chain reaction fingerprinting for the genotypic identification of mutants streptococci from humans[J]. Oral Microbiol Immunol, 1998,13(1):17.
  • 3Saarela M,Hannula J,Matto J,et al. Typing of mutants streptococci by arbitrarily primed polymerase chain reaction[J]. Archs Oral Biol, 1996,41(8/9):821~826.
  • 4Sambrook J,Fritsch EF,Maniatis T. 分子克隆[M]. 第2版. 北京:科学出版社, 1993.456~494.
  • 5Singer SJ,Tanguay RL,Riggs AD. Use of Chelex to improve the PCR signal from a small number of cells[J]. Amplification, 1989,3(1):11~16.
  • 6Giraffa G,Rossetti L,Neviani E. An evaluation of Chelex-based DNA purification protocols for the typing of lactic acid bacteria[J]. J Microbiol Methods, 2000,42(2):175~184.
  • 7Sepp R,Szabo I,Uda H,et al. Rapid techniques for DNA extraction from routinely processed archival tissue for use in PCR[J]. J Clin Pathol, 1994,47(3):318~323.
  • 8Walsh PS,Metzger DA,Higuchi R. Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material[J]. Biotechniques, 1991,10(4):506~513.
  • 9Mohlenhoff P,Muller L,Gorbushina AA,et al. Molecular approach to the characterisation of fungal communities: methods for DNA extraction,PCR amplification and DGGE analysis of painted art objects[J]. FEMS Microbiol Lett, 2001,195(2):169~173.
  • 10陈勖奇,中华内科杂志,1993年,32卷,655页

共引文献17

同被引文献255

引证文献21

二级引证文献161

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部