A non-radioactive method for inexpensive quantitative RT-PCR. |
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| Authors: | M Maggiolini O Donzé D Picard |
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| Institution: | Département de Biologie Cellulaire, Université de Genève, Switzerland. |
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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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