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PubMed · 10430034

A non-radioactive method for inexpensive quantitative RT-PCR.

M Maggiolini·O Donzé·D Picard
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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.

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Base SequenceBlotting, SouthernDNA PrimersDNA, ComplementaryDeoxyuracil NucleotidesDigoxigeninElectrophoresis, Agar GelHumansLuminescent MeasurementsRNA, MessengerReceptors, EstrogenReference StandardsReverse Transcriptase Polymerase Chain ReactionSensitivity and SpecificityTumor Cells, CulturedJournal ArticleResearch Support, Non-U.S. Gov't

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M Maggiolini, O Donzé, D Picard. 1999. A non-radioactive method for inexpensive quantitative RT-PCR.. https://doi.org/10.1515/bc.1999.086

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