4种古DNA抽提方法效果比较

doi:国家基础学科人才培养基金资助课题(J053018

• 专题研究-体质人类学 • 上一篇下一篇

4种古DNA抽提方法效果比较

蔡大伟^1,2，王海晶^2,3，韩璐²，李胜男²，周慧^1,2，朱泓¹

1.吉林大学边疆考古研究中心古DNA实验室，吉林长春130012；2. 吉林大学生命科学学院大分子实验室，吉林长春130023；3.吉林大学化学学院化学教学与研究中心，吉林长春130012

收稿日期:2006-02-21 修回日期:1900-01-01 出版日期:2007-01-28 发布日期:2007-01-28
通讯作者: 周慧

Comparisons of effects of four methods for ancient DNAextraction

CAI Da-wei^1,2, WANG Hai-jing²,3, HAN Lu², LI Sheng-nan²，ZHOU Hui^1,2，ZHU Hong¹

(1. Ancient DNA Laboratory, Research Center for Chinese Frontier Archaeology , Jilin University , Changchun 130012, China; 2.Macromolecular Laboratory,College of Life Science, Jilin University, Changchun 130023,China； 3. Teaching and Research Center of Chemistry,College of Chemistry, Jilin University, Changchun 130021, China

Received:2006-02-21 Revised:1900-01-01 Online:2007-01-28 Published:2007-01-28
Contact: ZHOU Hui

摘要/Abstract

摘要： 目的：寻找到一种简便、快速、可靠、有效的古DNA抽提方法。方法：选择基于硅粒法和硅离心柱法的四种抽提方法(方法A：Modified Silica Particles Method;方法B：Ceneclean Method;方法C：QIAamp Method;方法D：Modified QIAquick method))分别抽提5个距今约4 000年的古绵羊线粒体DNA (mtDNA)，并进行PCR扩增，通过琼脂糖凝胶电泳观察扩增成功与否，根据扩增成功率判断抽提方法的优劣。结果：4种方法的扩增成功率从20%～100%, 基于硅离心柱法的抽提效果 (60%，100%) 明显的优于基于硅粒法 (20%，40%)，其中Modified Silica Particles Method 的效果最差 (20%)，Modified QIAquick Method的效果最好 (100%)。　结论：Modified QIAquick Method方法使用Centricon○[KG-*4/5]〖HT7”〗R 超滤管浓缩DNA抽提裂解液，结合硅离心柱进行DNA纯化，能有效去除PCR抑制物，同时得到较好的古DNA模板。

关键词: DNA抽提, 硅粒, 硅离心柱

Abstract: To find a simple, rapid, reliable and effective method for ancient DNA extraction. Methods Four methods based on silica particles and silica-based spin columns(A:Modified Silica Particles Method;B:Geneclean Method;C:QIAamp Mothod;D:Modified QIAquick Method) were respectively used to extract mitochondrial DNA (mtDNA) from 5 ancient sheep dated to 4 000 years ago. PCR amplification of mtDNA was performed, and its results were evaluated by agarose gel electrophoresis. The effects of 4 kinds of methods mentioned above were determined by examining amplification success rates of DNA from 5 ancient sheep. Results The amplification success rates of 4 methods were different from 20% to 100%, and the effects of extraction based on silica-based spin columns (60%, 100%) were distinctly better than those based on silica particles (20%, 40%). The effect of extraction based on modified silica particles was the worst, and based on modified QIAquick was the best among 4 methods. Conclusion Modified QIAquick Method is a highly effective method, with a 100% success rate, in which Centricon○[KG-*4/5]〖WT7”〗R ultrafiltration devices are used to concentrate teeth digest solution and purify DNA through silica-based spin columns. It can effectively remove the inhibitor as well as obtain preferable DNA template.

Key words: DNA extraction, silica particles, silica-based spin columns

中图分类号:

Q751

蔡大伟，王海晶，韩璐，李胜男，周慧，朱泓. 4种古DNA抽提方法效果比较[J]. J4, 2007, 33(1): 13-16.

CAI Da-wei, WANG Hai-jing,3, HAN Lu, LI Sheng-nan，ZHOU Hui，ZHU Hong. Comparisons of effects of four methods for ancient DNAextraction[J]. J4, 2007, 33(1): 13-16.

4种古DNA抽提方法效果比较

Comparisons of effects of four methods for ancient DNAextraction

RichHTML

赞

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 0

Metrics

本文评价

推荐阅读 0