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Efficient reamplification of differential display products by transient ligation and thermal asymmetric PCR
S Bonnet1, G Prévot, C Bourgouin
1Unité d'Ecologie des Systèmes Vectoriels, Institut Pasteur, 25 rue du Docteur Roux, 75015 Paris, France.
Nucleic Acids Research
|March 21, 1998
Abstract:
A new method for specific reamplification of DDRT-PCR products is presented. After transient ligation of the primary DDRT-PCR fragments into a T-vector, the cDNAs of interest were reamplified by hemi-nested PCR and thermally asymmetric cycles. In contrast to the originally described protocol, this method of reamplification is specific, sensitive, reproducibly gives a high yield of DNA and allows direct sequencing of the reamplified product without purification or cloning.