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Scientific Article
unige:4656 
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A non-radioactive method for inexpensive quantitative RT-PCR |
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| Published in | Biological Chemistry. 1999, vol. 380, no. 6, p. 695-7 | |
| 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 | |
| Identifiers | DOI: 10.1515/BC.1999.086 PMID: 10430034 | |
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| 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. 695-7. doi: 10.1515/BC.1999.086 https://archive-ouverte.unige.ch/unige:4656 |
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