A non-radioactive method for inexpensive quantitative RT-PCR

Biol Chem. 1999 Jun;380(6):695-7. doi: 10.1515/BC.1999.086.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Blotting, Southern
  • DNA Primers
  • DNA, Complementary
  • Deoxyuracil Nucleotides
  • Digoxigenin / analogs & derivatives
  • Electrophoresis, Agar Gel
  • Humans
  • Luminescent Measurements
  • RNA, Messenger / analysis
  • RNA, Messenger / genetics
  • Receptors, Estrogen / genetics
  • Reference Standards
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Tumor Cells, Cultured


  • DNA Primers
  • DNA, Complementary
  • Deoxyuracil Nucleotides
  • RNA, Messenger
  • Receptors, Estrogen
  • digoxigenin-11-deoxyuridine triphosphate
  • Digoxigenin