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Full-Length Open Reading Frame Amplification of Hepatitis C Virus

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@article{a314aa4c97524f5b99ac35d13f1fb4c9,
title = "Full-Length Open Reading Frame Amplification of Hepatitis C Virus",
abstract = "The purpose of this method is to amplify the full coding sequence of hepatitis C virus (HCV) by a single round reverse transcriptase-polymerase chain reaction (RT-PCR) approach. Our method relies on a highly robust and sensitive RNA extraction procedure and cutting-edge RT-PCR enzymes, all of which have been rigorously tested and optimized. This will not only allow for robust amplification of the entire open reading frame (ORF) of HCV for sequencing by Sanger or next-generation sequencing (NGS), but can also be used for cloning of the ORF of uncharacterized samples and for linkage analysis of mutations on individual genomes spanning the entire ORF. The method has been validated on a variety of samples, including sera from HCV patients and cell-culture supernatants.",
keywords = "Cloning, Full-length ORF RT-PCR, HCV, Hepatitis C virus, Long PCR, Next-generation sequencing, RNA extraction, Sanger sequencing",
author = "Ulrik Fahn{\o}e and Jens Bukh",
year = "2019",
doi = "10.1007/978-1-4939-8976-8_5",
language = "English",
volume = "1911",
pages = "85--91",
journal = "Methods in Molecular Biology",
issn = "1064-3745",
publisher = "Humana Press, Inc",

}

RIS

TY - JOUR

T1 - Full-Length Open Reading Frame Amplification of Hepatitis C Virus

AU - Fahnøe, Ulrik

AU - Bukh, Jens

PY - 2019

Y1 - 2019

N2 - The purpose of this method is to amplify the full coding sequence of hepatitis C virus (HCV) by a single round reverse transcriptase-polymerase chain reaction (RT-PCR) approach. Our method relies on a highly robust and sensitive RNA extraction procedure and cutting-edge RT-PCR enzymes, all of which have been rigorously tested and optimized. This will not only allow for robust amplification of the entire open reading frame (ORF) of HCV for sequencing by Sanger or next-generation sequencing (NGS), but can also be used for cloning of the ORF of uncharacterized samples and for linkage analysis of mutations on individual genomes spanning the entire ORF. The method has been validated on a variety of samples, including sera from HCV patients and cell-culture supernatants.

AB - The purpose of this method is to amplify the full coding sequence of hepatitis C virus (HCV) by a single round reverse transcriptase-polymerase chain reaction (RT-PCR) approach. Our method relies on a highly robust and sensitive RNA extraction procedure and cutting-edge RT-PCR enzymes, all of which have been rigorously tested and optimized. This will not only allow for robust amplification of the entire open reading frame (ORF) of HCV for sequencing by Sanger or next-generation sequencing (NGS), but can also be used for cloning of the ORF of uncharacterized samples and for linkage analysis of mutations on individual genomes spanning the entire ORF. The method has been validated on a variety of samples, including sera from HCV patients and cell-culture supernatants.

KW - Cloning

KW - Full-length ORF RT-PCR

KW - HCV

KW - Hepatitis C virus

KW - Long PCR

KW - Next-generation sequencing

KW - RNA extraction

KW - Sanger sequencing

UR - http://www.scopus.com/inward/record.url?scp=85059244974&partnerID=8YFLogxK

U2 - 10.1007/978-1-4939-8976-8_5

DO - 10.1007/978-1-4939-8976-8_5

M3 - Journal article

VL - 1911

SP - 85

EP - 91

JO - Methods in Molecular Biology

JF - Methods in Molecular Biology

SN - 1064-3745

ER -

ID: 56118500