Full-length open reading frame amplification of hepatitis C virus
Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-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.
|Title of host publication||Hepatitis C Virus Protocols|
|Number of pages||7|
|Publication status||Published - 2019|
|Series||Methods in Molecular Biology|
- Cloning, Full-length ORF RT-PCR, HCV, Hepatitis C virus, Long PCR, Next-generation sequencing, RNA extraction, Sanger sequencing