Full-length open reading frame amplification of hepatitis C virus
Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review
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Full-length open reading frame amplification of hepatitis C virus. / Fahnøe, Ulrik; Bukh, Jens.
Hepatitis C Virus Protocols. Humana Press, 2019. p. 85-91 (Methods in Molecular Biology, Vol. 1911).Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review
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TY - CHAP
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
U2 - 10.1007/978-1-4939-8976-8_5
DO - 10.1007/978-1-4939-8976-8_5
M3 - Book chapter
C2 - 30593619
AN - SCOPUS:85059244974
SN - 978-1-4939-8975-1
T3 - Methods in Molecular Biology
SP - 85
EP - 91
BT - Hepatitis C Virus Protocols
PB - Humana Press
ER -
ID: 212851632