Methods of DNA extraction from bacteria for PCR and Real-time PCR

HU Xiao-hong; et al

您当前的位置：

首页 >

文章列表页 >

Methods of DNA extraction from bacteria for PCR and Real-time PCR

更新时间：2024-01-17

- Methods of DNA extraction from bacteria for PCR and Real-time PCR
- Journal of Chongqing Medical University Issue 2, Pages: 155-158(2008)
- 作者机构：
  
  1. 重庆医科大学基础医学院细胞生物学及遗传学教研室
  2. 第三军医大学分子遗传学教研室
- 作者简介：
- 基金信息：
- DOI：
  CLC： R346
- Published：2008，
扫描看全文
[1]胡晓红,彭惠民,刘昕,黄正根,袁科.PCR及Real-time PCR评价细菌DNA提取方法[J].重庆医科大学学报,2008(02):155-158.

HU Xiao-hong, et al. Methods of DNA extraction from bacteria for PCR and Real-time PCR. [J]. Journal of Chongqing Medical University (2):155-158(2008)
[1]胡晓红,彭惠民,刘昕,黄正根,袁科.PCR及Real-time PCR评价细菌DNA提取方法[J].重庆医科大学学报,2008(02):155-158. DOI：

HU Xiao-hong, et al. Methods of DNA extraction from bacteria for PCR and Real-time PCR. [J]. Journal of Chongqing Medical University (2):155-158(2008) DOI：

摘要

目的:比较几种不同的细菌DNA提取方法

建立一种适于PCR和Real-time PCR的细菌DNA提取方法。方法:分别用蛋白酶K法、碱裂解法、Chelex-100+NP40、Chelex-100+TritonX-100法和水煮法提取同种浓度大肠杆菌DNA

测定OD260/280;用不同方法提取不同浓度的大肠杆菌DNA

进行PCR和实时荧光定量PCR扩增

比较灵敏度。结果:5种方法提取细菌DNA

其中Chelex-100+NP40法纯度（OD260/280=1.79±0.03）最高

此方法所提取的DNA产物进行PCR及Real-time PCR扩增未见假阳性及假阴性

其灵敏度为10个/ml细菌浓度。结论:Chelex-100+NP40的细菌DNA提取方法纯度及灵敏度高

且适应于PCR及Real-time PCR。

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.

关键词

DNA提取PCR实时荧光定量PCR

Keywords

DNA extractionPCRreal-time PCR

references

Views

2567

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Analysis of pertussis and pertussis syndrome

Analyze the associativity between Helicobacter pylori infection and cholecystolithiasis

Relationship Between A1675G Polymorphism of Angiotensin II type 2 - receptor Gene and Essential Hypertension

Construction and expression of Retroviral Vector containing HBx gene

Establishment and application of a fluorescent quantitative pcr method for cccDNA of HBV

Related Author

Ye Zehui

Huang Ying

Yan Li

谢健

ZHU Yujuan

周卫东

肖思洁

杨华

Related Institution

Department of Respiratory Medicine,the Children’s Hospital,Chongqing Medical University

成都三六三医院消化内科四川省卫生干部管理学院四川大学华西医院

四川大学华西医院

四川省卫生干部管理学院

Department of Gastroenterology, Hospital of Chengdu

⁰