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The Capital Region of Denmark - a part of Copenhagen University Hospital
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Full-Length Open Reading Frame Amplification of Hepatitis C Virus

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  1. Determination of Binding Kinetics of Intrinsically Disordered Proteins by Surface Plasmon Resonance

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  2. Quantity and Quality of Basophil RNA Depend on the RNA Extraction Technique

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  3. Analysis of Mass Cytometry Data

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  4. Assessment of Peptidylarginine Deiminase Activity by ELISA Using Human Fibrinogen as Substrate

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  5. In Vitro Neutralization Assay Using Cultured Hepatitis C Virus

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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
JournalMethods in molecular biology
Volume1911
Pages (from-to)85-91
Number of pages7
ISSN1064-3745
DOIs
Publication statusPublished - 2019

    Research areas

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

ID: 56118500