A non-radioactive method for inexpensive quantitative RT-PCR

被引：35
|
作者：
Maggiolini, M [1 ]
Donzé, O [1 ]
Picard, D [1 ]
机构：
[1] Univ Geneva, Dept Biol Cellulaire, CH-1211 Geneva 4, Switzerland
来源：
BIOLOGICAL CHEMISTRY | 1999年 / 380卷 / 06期
关键词：
breast cancer cells; cDNA; chemiluminescence; digoxigenin; estrogen receptor; gene expression;
D O I：
10.1515/BC.1999.086
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
We present a novel method for quantitative RT-PCR that involves direct incorporation of digoxigenin-11-dUTP (DIG-dUTP) during amplification of cDNAs, separation of RT-PCR products by agarose gel electrophoresis, Southern transfer to a nylon membrane, and chemiluminescent detection with an anti-DIG antibody. The whole procedure can be done in about a day and has the following advantages: It is highly sensitive, specificity is confirmed by monitoring the size of the RT-PCR product, it is non-radioactive, quantitative, and does not require expensive specialized equipment.
引用
收藏
页码：695 / 697
页数：3
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