A simple method for the isolation of genomic DNA from mouse tail free of real-time PCR inhibitors

被引:31
作者
Burkhart, CA [1 ]
Norris, MD [1 ]
Haber, M [1 ]
机构
[1] Childrens Canc Inst Australia Med Res, Sydney, NSW 2031, Australia
来源
JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS | 2002年 / 52卷 / 02期
基金
英国医学研究理事会;
关键词
real-time PCR; inhibitors; mouse tail; genomic DNA;
D O I
10.1016/S0165-022X(02)00052-0
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Although real-time PCR is a rapid, quantitative method for the analysis of gene and RNA levels, the presence of inhibitors in samples is an obstacle to its successful use. We have found that genomic DNA isolated from mouse tail tips using a standard proteinase K digestion method caused marked inhibition of real-time PCR. Inhibition was specific for mouse tail DNA since genomic DNA isolated from other tissue sources using the same methodology was readily amplified. We have therefore developed a nonproteinase K DNA isolation method involving the use of Chelex((R)) 100 resin. This method produces mouse tail DNA that is free of real-time PCR inhibitors. (C) 2002 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:145 / 149
页数:5
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