Comparison of three methods for total DNA extraction from mouse models
CSTR:
Author:
  • Article
  • | |
  • Metrics
  • |
  • Reference [12]
  • | | | |
  • Comments
    Abstract:

    Objective To establish a simple, fast and economic total DNA extraction method to serve the rapid identification of model mouse genotype in large number of mice. Methods Three methods, i.e. phenol extraction, isopropyl alcohol precipitation and mouse ear boiling methods were used to extract the total DNA from ten C57-rasmodel mice. The purity and yield of DNA obtained by the three methods were compared, and polymerase chain reaction (PCR) assay was used to compare the efficacy of the three extraction methods. Results Among the three methods, phenol extraction was the best and isopropyl alcohol precipitation was the poorest in DNA yield. In terms of DNA purity, the phenol extraction was the best and the mouse ear boiling method was the poorest. All the three methods could be used to extract the total DNA from mice serving as template of PCR reaction for the mouse genotype identification. The time consumption of three methods are 12.5 hr,13 hr and 0.18 hr. Mouse ear boiling method was significantly lower than that of the other two methods (P< 0.01 ),.The obtained total DNA can be stored at conventional-20℃ for 7 days and 30 days later still can be used as a template for PCR reaction. Conclusions Among the three methods studied, the mouse ear boiling method is simple and with the lowest cost, so it is feasible for total DNA extraction in scaled genotyping experiments.

    Reference
    [1] 卢一凡,田靫,邓继先. 转基因动物鉴定技术的研究进展[J].生物工程进展, 2000,20(3):60-61.
    [2] 赵彦青, 纪香,赵宝成. 转基因动物的研究与应用及其存在问题[C].“科技进步推进畜牧业现代化”科技论文集, 2011年.
    [3] 杨继山, 潘庆杰, 董晓. 转基因动物检测方法的研究进展[J].中国农业科技导报, 2010, 12(3):45-49.
    [4] 萨姆布鲁克J, 拉塞尔DW,主编, 黄培堂译.分子克隆实验指南 (第三版)[M]. 北京: 科学出版社, 2002, 487-509.
    [5] Linz U,Delling U,Rubsamen-Waigmann H. Systematic studies on parameters influencing the performance of the polymerase chainreaction[J]. J ClinChem Biochem.1990,28(1):5-13.
    [6] 任亮, 苏玉虹,巴彩凤,等. PCR引物设计技巧[J]. 现代畜牧兽医,2005,6:49
    [7] 卢圣栋. 现代分子生物学实验技术[M]. 北京: 高等教出版社, 1993:96.
    [8] Abbot C, Prove S, Vivian N, et al. PCR as a rapid screening method for transgenic mouse[J]. Trends in Genetics,1988,4(11):325.
    [9] 李永明,赵玉琪. 实用分子生物学方法手册[M]北京: 科学出版社, 1999, 21-23.
    [10] 肖波,李岩, 屈慧歌,等. 两种动物基因组DNA 提取方法的比较[J].烟台师范学院学报(自然科学版),2005, 21(1):56-58.
    [11] 杨建雄. 生物化学与分子生物学实验技术教程[M]. 北京: 科学出版社, 2002. 71-131
    [12] 鲍毅新, 孙波, 张龙龙, 等. 对动物组织DNA提取方法的改进及PCR检测[J].浙江师范大学学报(自然科学版), 2009, 32(3):318-320.
    Related
    Cited by
    Comments
    Comments
    分享到微博
    Submit
Get Citation
Share
Article Metrics
  • Abstract:2052
  • PDF: 1606
  • HTML: 0
  • Cited by: 0
History
  • Revised:May 13,2014
  • Online: August 19,2014
Article QR Code