Methods of DNA extraction from bacteria for PCR and Real-time PCR
- VernacularTitle:PCR及Real-time PCR评价细菌DNA提取方法
- Author:
Xiaohong HU
- Publication Type:Journal Article
- Keywords:
DNA extraction;
PCR;
Real-time PCR
- From:
Journal of Chongqing Medical University
1986;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To compare the different DNA extraction methods and establish an efficient and sensitive method of DNA extraction from bacteria for PCR and Real-time PCR.Methods:Bacillus coli DNA with the same concentration was extracted by 5 different DNA extraction methods,the Proteinase K method,the alkali-lysis method,the Chelex-100 with NP40 or Triton X-100 and the water-boiling method to determine OD260/280.Bacillus coli DNA with different concentrations wewe extracted to be amplified by PCR and Real-time PCR for compariing the sensitivity.Results:Among these extraction methods,Chelex-100 + NP40 method was with the best purity and sensitivity and offered a higher OD260/280(1.79?0.03).Neither false positive nor false negative was found in PCR and Real-time PCR,The sensitivity was at 10/ml of bacteria concentratio.Conclusion:The DNA extraction based on Chelex-100 + NP40 is effective,specific and sensitive,and it accommodate to PCR and Real-time PCR.
