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
相关论文
共 50 条
  • [31] PCR-Mediated synthesis of exogenous competitors for quantitative RT-PCR
    Schneeberger, C
    Zeillinger, R
    BIOTECHNIQUES, 1996, 20 (03) : 360 - &
  • [32] A new quantitative RT-PCR assay for the vasoconstrictor endothelin
    Lederer, EB
    Bastani, K
    Goebel, FD
    RESEARCH IN EXPERIMENTAL MEDICINE, 1996, 196 (02) : 77 - 86
  • [33] Detection and quantitation of bovine respiratory syncytial virus using real-time quantitative RT-PCR and quantitative competitive RT-PCR assays
    Achenbach, JE
    Topliff, CL
    Vassilev, VB
    Donis, RO
    Eskridge, KM
    Kelling, CL
    JOURNAL OF VIROLOGICAL METHODS, 2004, 121 (01) : 1 - 6
  • [34] A modified PCR-based method for rapid non-radioactive detection of clinically important pathogens
    Gorelov, VN
    Dumon, K
    Barteneva, NS
    Roher, HD
    Goretzki, PE
    MICROBIOLOGY AND IMMUNOLOGY, 1996, 40 (09) : 611 - 616
  • [35] A HIGHLY SENSITIVE, ONE-STEP QUANTITATIVE RT-PCR METHOD FOR THE DETECTION OF EGFRVIII
    Li, Gordon
    Wong, Albert
    NEURO-ONCOLOGY, 2008, 10 (05) : 761 - 761
  • [36] Establishment and application of quadruple fluorescence quantitative RT-PCR method for the identification of waterfowl astrovirus
    Li, Yang
    Luo, Juan
    Zhang, Fuyou
    Shang, Jiajing
    Deng, Chunran
    Feng, Yingjie
    Meng, Ge
    Jiang, Wenming
    Yu, Xiaohui
    Liu, Guanhui
    Liu, Hualei
    FRONTIERS IN MICROBIOLOGY, 2024, 15
  • [37] Pre-amplification as a method for improvement of quantitative RT-PCR analysis of circulating miRNAs
    Sekovanic, Ankica
    Dorotic, Adrijana
    Jurasovic, Jasna
    Pasalic, Daria
    Kovacic, Jelena
    Stasenko, Sandra
    Mioc, Tatjana
    Piasek, Martina
    Orct, Tatjana
    BIOCHEMIA MEDICA, 2021, 31 (01) : 1 - 6
  • [38] DEVELOPMENT OF A QUANTITATIVE REAL TIME RT-PCR METHOD FOR BRCA1 mRNA
    Kroupis, C.
    Stathopoulou, A.
    Malamos, N.
    Georgoulias, V.
    Lianidou, E.
    ANTICANCER RESEARCH, 2004, 24 (5D) : 3542 - 3542
  • [39] A new quantitative RT-PCR method for sensitive detection of dengue virus in serum samples
    Sadon, Nadine
    Delers, Anne
    Jarman, Richard G.
    Klungthong, Chonticha
    Nisalak, Ananda
    Gibbons, Robert V.
    Vassilev, Ventzislav
    JOURNAL OF VIROLOGICAL METHODS, 2008, 153 (01) : 1 - 6
  • [40] Quantification of tissue factor gene expression using fluorogenic quantitative RT-PCR method
    Guan, Ming
    Su, Bing
    Lu, Yuan
    Fudan University Journal of Medical Sciences, 2002, 29 (01): : 66 - 68
12345