Detection of G1691A mutation in factor V in human and Fingerprinting of HIV-1 quasispecies. (a) Schematic representation of 611 bp DNA fragment of factor V indicating G1691A mutation associated with venous thrombosis. The red and green dots indicate the position of IRD700 and IRD800-labelled primers, respectively. (b) Genomic DNA was extracted from leg hairs of a non-carrying (lane 3 and 4) and heterozygous individual for the G1691A mutation (lane 1 and 2). The 611 bp DNA fragment corresponding to the region harboring the mutation was PCR amplified, denaturated/renaturated prior to digestion with ENDO1 and analyzed on on LI-COR4300 DNA Analyzer. Sizes of the digested fragments are indicated on the left. (c) Chromatogram of the sequence of the human Factor V fragment from heterozygous individual showing the G1691A mutation. (d) Genomic DNAs were extracted from peripheral blood mononuclear cells of an HIV-1 infected patient before (lane 1) and after 48 weeks of antiretroviral therapy (lane 2). DNA fragment of 843 bp coding for the reverse transcriptase gene of HIV-1 was PCR amplified, denaturated/renaturated prior to digestion with ENDO1 and analyzed on LI-COR4300 DNA Analyzer. The collected image from both channels is shown (IRD700 and IRD800 channels). Examples of bands of invariant intensity that indicate stable mutations are indicated by filled arrow. Examples of bands indicating mutations that either appeared or were lost upon treatment are indicated by empty arrows.