אסיף מאגר המחקר החקלאי

A procedure to amplify cDNA from dsRNA templates using the polymerase chain reaction

Year:

1991

Source of publication :

Authors :

Volume :

2

Co-Authors:

Wexler, A., S. Tolkowsky Laboratory, A.R.O. The Volcani Center, Bet Dagan 20250, Israel
Mawassi, M., S. Tolkowsky Laboratory, A.R.O. The Volcani Center, Bet Dagan 20250, Israel
Lachman, O., S. Tolkowsky Laboratory, A.R.O. The Volcani Center, Bet Dagan 20250, Israel
Amit, B., S. Tolkowsky Laboratory, A.R.O. The Volcani Center, Bet Dagan 20250, Israel
Wortzel, A., S. Tolkowsky Laboratory, A.R.O. The Volcani Center, Bet Dagan 20250, Israel
Bar-Joseph, M., S. Tolkowsky Laboratory, A.R.O. The Volcani Center, Bet Dagan 20250, Israel

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Abstract:

We have devised a method to amplify complementary DNA (cDNA) derived from double-stranded RNA (dsRNA) templates to which a 20-mer polylinker oligodeoxynucleotide (oA), was enzymatically ligated. A second oligonucleotide (oB) complementary to oA was used as a primer for the synthesis of cDNA on the chimeric RNA-oA molecules. The cDNAs synthesized on the two RNA strands were annealed and extended using Taq polymerase and the DNA was amplified by PCR using oB as the primer. The resulting DNA was restricted enzymatically and cloned in a pBluescript vector. We have used this method to develop a range of cDNA clones from the dsRNAs of two plant viruses, cucumber mosaic virus and citrus tristeza virus.

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