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Methods for viral RNA isolation and PCR amplification for sequencing of near full-length HIV-1 genomes
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SYSNO ASEP 0317965 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Ostatní články Title Methods for viral RNA isolation and PCR amplification for sequencing of near full-length HIV-1 genomes Title Metody RNA isolace a PCR amplifikace pro sekvenaci téměř celých genomů HIV-1 Author(s) Kemal, K.S. (US)
Reiniš, Milan (UMG-J) RID
Weiser, B. (US)
Burger, H. (US)Source Title The Nucleus, Volume 1: Nuclei and Subnuclear Components. - Totowa : Humana Press, 2008 / Hancock Ronald - ISBN 978-1-58829-977-2
Roč. 485, - (2009), s. 3-14Number of pages 12 s. Language eng - English Country US - United States Keywords HIV-1 viral RNA ; HIV-1 primers ; long RT-PCR Subject RIV EB - Genetics ; Molecular Biology CEZ AV0Z50520514 - UMG-J (2005-2011) Annotation 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. Workplace Institute of Molecular Genetics Contact Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Year of Publishing 2009
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