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A procedure to amplify cDNA from dsRNA templates using the polymerase chain reaction
Year:
1991
Authors :
Bar-Joseph, Moshe
;
.
Lachman, Oded
;
.
Mawassi, Munir
;
.
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
Facilitators :
From page:
273
To page:
279
(
Total pages:
7
)
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.
Note:
Related Files :
article
Citrus tristeza virus
Cucumber mosaic virus
Cucumis sativus
double stranded RNA
gene amplification
Polymerase Chain Reaction
Show More
Related Content
More details
DOI :
Article number:
0
Affiliations:
Database:
Scopus
Publication Type:
article
;
.
Language:
English
Editors' remarks:
ID:
19541
Last updated date:
02/03/2022 17:27
Creation date:
16/04/2018 23:29
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Scientific Publication
A procedure to amplify cDNA from dsRNA templates using the polymerase chain reaction
2
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
A procedure to amplify cDNA from dsRNA templates using the polymerase chain reaction
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.
Scientific Publication
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