Novel application of Phi29 DNA polymerase: RNA detection and analysis in vitro and in situ by target RNA-primed RCA.
RNA
; 15(5): 765-71, 2009 May.
Article
in En
| MEDLINE
| ID: mdl-19244362
We present a novel Phi29 DNA polymerase application in RCA-based target RNA detection and analysis. The 3'-->5' RNase activity of Phi29 DNA polymerase converts target RNA into a primer and the polymerase uses this newly generated primer for RCA initiation. Therefore, using target RNA-primed RCA, padlock probes may be targeted to inner RNA sequences and their peculiarities can be analyzed directly. We demonstrate that the exoribonucleolytic activity of Phi29 DNA polymerase can be successfully applied in vitro and in situ. These findings expand the potential for detection and analysis of RNA sequences distanced from 3'-end.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
RNA
/
Genetic Techniques
/
Bacillus Phages
/
DNA-Directed DNA Polymerase
Type of study:
Diagnostic_studies
Limits:
Humans
Language:
En
Journal:
RNA
Journal subject:
BIOLOGIA MOLECULAR
Year:
2009
Document type:
Article
Affiliation country:
Lithuania
Country of publication:
United States