Full-length open reading frame amplification of hepatitis C virus

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearchpeer-review

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.

Original languageEnglish
Title of host publicationHepatitis C Virus Protocols
Number of pages7
PublisherHumana Press
Publication date2019
ISBN (Print)978-1-4939-8975-1
ISBN (Electronic)978-1-4939-8976-8
Publication statusPublished - 2019
SeriesMethods in Molecular Biology

    Research areas

  • Cloning, Full-length ORF RT-PCR, HCV, Hepatitis C virus, Long PCR, Next-generation sequencing, RNA extraction, Sanger sequencing

ID: 212851632