HIV/SIV Single Genome Amplification and Sequencing
ACVP-05
- 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.
