title: Novel application of Phi29 DNA polymerase: RNA detection and analysis in vitro and in situ by target RNA-primed RCA.
abstract: 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.
authors.affiliation: Fermentas UAB, Vilnius LT-02241, Lithuania. laguna@fermentas.lt
authors.forename: Arunas
authors.initials: A
authors.lastname: Lagunavicius
authors.forename: Egle
authors.initials: E
authors.lastname: Merkiene
authors.forename: Zivile
authors.initials: Z
authors.lastname: Kiveryte
authors.forename: Agne
authors.initials: A
authors.lastname: Savaneviciute
authors.forename: Vilma
authors.initials: V
authors.lastname: Zimbaite-Ruskuliene
authors.forename: Tomas
authors.initials: T
authors.lastname: Radzvilavicius
authors.forename: Arvydas
authors.initials: A
authors.lastname: Janulaitis
mesh_terms: D017103:Bacillus Phages / Q000201:enzymology*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D005821:Genetic Techniques; D006367:HeLa Cells; D006801:Humans; D009711:Nucleotides / Q000378:metabolism; D012313:RNA / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D009711:Nucleotides; C034477:RNA primers; D012313:RNA; D004259:DNA-Directed DNA Polymerase