doi: 10.1515/BC.1999.086.
A non-radioactive method for inexpensive quantitative RT-PCR
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- PMID: 10430034
- DOI: 10.1515/BC.1999.086
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A non-radioactive method for inexpensive quantitative RT-PCR
Biol Chem.
1999 Jun.
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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