Amplification and fusion of long fragments of hepatitis C virus genome.
The 'long PCR' was used for amplification of hepatitis C virus (HCV) subgenomic fragments from liver. After testing several commercially available systems, it was found that Tth as the major enzyme is superior to using Taq. Employing a mixture of Tth and Vent polymerase (rTth polymerase, XL, Perkin Elmer) it was possible to amplify 4.6-kb and 9-kb fragments from biological samples containing as little as 10(2) and 10(4) viral copies, respectively. It was also demonstrated that 'long PCR' is useful for joining together large size amplification products.
Wang
L F
LF
Division of Transplantation Medicine, Pittsburgh Transplantation Institute, University of Pittsburgh Medical Center, PA 15213, USA.
Radkowski
M
M
Vargas
H
H
Rakela
J
J
Laskus
T
T
D019677:Artificial Gene Fusion / Q000379:methods*; D001482:Base Composition; D018076:DNA, Complementary / Q000302:isolation & purification; D016679:Genome, Viral; D016174:Hepacivirus / Q000235:genetics*; D006801:Humans; D008099:Liver / Q000821:virology; D016031:Liver Transplantation; D009924:Organ Culture Techniques; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000302:isolation & purification; D012194:RNA-Directed DNA Polymerase; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D013698:Templates, Genetic
D018076:DNA, Complementary; D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic; D012194:RNA-Directed DNA Polymerase