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

页码：695 / 697

页数：3

共 11 条

[1] Comparison of detection techniques for cytokine reverse transcriptase polymerase chain reaction; digoxigenin-labeled polymerase chain reaction permits sensitive detection of cytokine mRNA in rat heart allografts [J].