HIV/SIV Single Genome Amplification and Sequencing

  • Provides innovative sequencing techniques, molecular cloning, and viral evolution analysis. 
  • Identifies genetic changes in viral populations. 

The process: 

  • The Viral Evolution Core extracts viral RNA/DNA from cell-free or cell-associated samples. 
  • Complementary cDNA is generated as needed, and cDNA or DNA is diluted to a single copy prior to nested polymerase chain reaction (PCR) spanning the viral gene of interest. 
  • Following PCR, amplicons are selected by gel electrophoresis and sequenced using Sanger sequencing. 
    • Samples are diluted down to a single template copy per PCR reaction as calculated by Poisson probability distribution
  • Confirmation of single-molecule polymerase chain reaction is obtained by directly sequencing PCR products and excluding amplicons with evidence of more than one template. 
  • Provides comprehensive sequence analysis, including enumerating transmitted and founder genomes, phylogenetic tree construction, and genetic comparisons of diversity. 
  • Provides relevant data electronically, including a report with interpretations and conclusions. 

Includes sufficient basic consultation with a subject matter expert.