en
Scientific article
English

A non-radioactive method for inexpensive quantitative RT-PCR

Published inBiological chemistry, vol. 380, no. 6, p. 695-697
Publication date1999
Abstract

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.

Keywords
  • Base Sequence
  • Blotting, Southern
  • Chemiluminescent Measurements
  • DNA Primers
  • DNA, Complementary
  • Deoxyuracil Nucleotides
  • Digoxigenin/analogs & derivatives
  • Electrophoresis, Agar Gel
  • Humans
  • RNA, Messenger/analysis/genetics
  • Receptors, Estrogen/genetics
  • Reference Standards
  • Reverse Transcriptase Polymerase Chain Reaction/methods
  • Sensitivity and Specificity
  • Tumor Cells, Cultured
Citation (ISO format)
MAGGIOLINI, Marcello, DONZE, Olivier, PICARD, Didier. A non-radioactive method for inexpensive quantitative RT-PCR. In: Biological chemistry, 1999, vol. 380, n° 6, p. 695–697. doi: 10.1515/BC.1999.086
Main files (1)
Article (Published version)
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Identifiers
ISSN of the journal1431-6730
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