About: Methods for viral RNA isolation and PCR amplification for sequencing of near full-length HIV-1 genomes

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rdf:type	skos:Concept http://linked.opendata.cz/ontology/domain/vavai/Vysledek
Description	HIV-1 se replikuje v plasmě v průběhu virové infekce. Studia RNA z plasmy a dalších tělních tekutin proto reflektuje varianty viru v reálném čase. K získání genomových sekvencí odvozených z virové RNA je nutno nejprve amplifikovat RNA. V tomto článku jsou popsány metody RNA extrakce, její reversní transkripce na cDNA a amplifikace dlouhých DNA fragmentů za použití sady specifických primerů tak, aby vznikaly vzájemně se překrývající fragmenty DNA. Primery byly odvozeny ze sekvencí HIV-1 subtypu a metodika optimalizována na isolaci vzorků z plasmy. Nicméně protokol může být snadno upraven modifikacím použitých primerů na studium dalších HIV-1 subtypů. (cs) HIV-1 in plasma represents the viral quasispecies replicating in the patient at any given time. Studies of HIV-1 viral RNA from plasma or other body fluids therefore reflect the virus present in real time. To obtain near full-length genomic sequences derived from virion RNA it is first necessary to carefully isolate and amplify the RNA.The procedure described below, involves viral RNA extraction, reverse transcription (RT) of the extracted RNA to produce cDNA copies, and PCR amplification of long HIV-1 gene fragments using site-specific, overlapping primers. The primers are based on subtype B HIV-1 strains, and plasma specimens are used in the procedures. However, the protocol can easily be adapted to other HIV-1 subtypes by modifying the primers to match the subtype of interest. HIV-1 in plasma represents the viral quasispecies replicating in the patient at any given time. Studies of HIV-1 viral RNA from plasma or other body fluids therefore reflect the virus present in real time. To obtain near full-length genomic sequences derived from virion RNA it is first necessary to carefully isolate and amplify the RNA.The procedure described below, involves viral RNA extraction, reverse transcription (RT) of the extracted RNA to produce cDNA copies, and PCR amplification of long HIV-1 gene fragments using site-specific, overlapping primers. The primers are based on subtype B HIV-1 strains, and plasma specimens are used in the procedures. However, the protocol can easily be adapted to other HIV-1 subtypes by modifying the primers to match the subtype of interest. (en)
Title	Methods for viral RNA isolation and PCR amplification for sequencing of near full-length HIV-1 genomes Methods for viral RNA isolation and PCR amplification for sequencing of near full-length HIV-1 genomes (en) Metody RNA isolace a PCR amplifikace pro sekvenaci téměř celých genomů HIV-1 (cs)
skos:prefLabel	Methods for viral RNA isolation and PCR amplification for sequencing of near full-length HIV-1 genomes Methods for viral RNA isolation and PCR amplification for sequencing of near full-length HIV-1 genomes (en) Metody RNA isolace a PCR amplifikace pro sekvenaci téměř celých genomů HIV-1 (cs)
skos:notation	RIV/68378050:_____/09:00317965!RIV09-AV0-68378050
http://linked.open...avai/riv/aktivita	Z
http://linked.open...avai/riv/aktivity	Z(AV0Z50520514)
http://linked.open...iv/cisloPeriodika	-
http://linked.open...vai/riv/dodaniDat	2009
http://linked.open...aciTvurceVysledku	Reiniš, Milan
http://linked.open.../riv/druhVysledku	J - Článek v odborném periodiku
http://linked.open...iv/duvernostUdaju	S - Úplné a pravdivé údaje nepodléhající ochraně podle zvláštních právních předpisů
http://linked.open...titaPredkladatele	Ústav molekulární genetiky AV ČR, v. v. i.
http://linked.open...dnocenehoVysledku	325709
http://linked.open...ai/riv/idVysledku	RIV/68378050:_____/09:00317965
http://linked.open...riv/jazykVysledku	eng - angličtina
http://linked.open.../riv/klicovaSlova	HIV-1 viral RNA; HIV-1 primers; long RT-PCR (en)
http://linked.open.../riv/klicoveSlovo	HIV-1 primers HIV-1 viral RNA long RT-PCR
http://linked.open...odStatuVydavatele	US - Spojené státy americké
http://linked.open...ontrolniKodProRIV	[F581626ABF79]
http://linked.open...i/riv/nazevZdroje	The Nucleus, Volume 1: Nuclei and Subnuclear Components
http://linked.open...in/vavai/riv/obor	EB
http://linked.open...ichTvurcuVysledku	1 (xsd:int)
http://linked.open...cetTvurcuVysledku	4 (xsd:int)
http://linked.open...UplatneniVysledku	2009
http://linked.open...v/svazekPeriodika	485
http://linked.open...iv/tvurceVysledku	Reiniš, Milan Burger, H. Kemal, K. S. Weiser, B.
http://linked.open...n/vavai/riv/zamer	Molekulárně genetické a buněčné základy klíčových biologických procesů: genová exprese, onkogeneze, replikace virů, imunita a vývoj organismů
issn	1064-3745
number of pages	12 (xsd:int)
is http://linked.open...avai/riv/vysledek of	Methods for viral RNA isolation and PCR amplification for sequencing of near full-length HIV-1 genomes Methods for viral RNA isolation and PCR amplification for sequencing of near full-length HIV-1 genomes Methods for viral RNA isolation and PCR amplification for sequencing of near full-length HIV-1 genomes Methods for viral RNA isolation and PCR amplification for sequencing of near full-length HIV-1 genomes

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