A Non-Radioactive Method for Inexpensive Quantitative RT-PCR
- 1 June 1999
- journal article
- research article
- Published by Walter de Gruyter GmbH in Biological Chemistry
- Vol. 380 (6), 695-697
- https://doi.org/10.1515/bc.1999.086
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
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