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A non-radioactive method for inexpensive quantitative RT-PCR

Maggiolini, Marcello
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 SequenceBlotting, SouthernChemiluminescent MeasurementsDNA PrimersDNA, ComplementaryDeoxyuracil NucleotidesDigoxigenin/analogs & derivativesElectrophoresis, Agar GelHumansRNA, Messenger/analysis/geneticsReceptors, Estrogen/geneticsReference StandardsReverse Transcriptase Polymerase Chain Reaction/methodsSensitivity and SpecificityTumor Cells, Cultured
PMID: 10430034
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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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Deposited on : 2009-12-10

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