Fig. 1

Electrophoretic mobility patterns of PCR products from HCV detection, genotyping and (pre)cloning reactions In all gels first lane displays the molecular weight marker - 100 bp in ‘A’ and ‘B’ and 1 kb in ‘C’ and ‘D’. (a) HCV RNA detection in plasma samples. Lane 2 shows cDNA band (268 bp) of globin mRNA used as HCV positive control. Lanes 4 and 5 show 268 bp product indicating presence of HCV RNA in corresponding samples, HCV 152 and HCV 173. (b) Primer-specific genotyping of HCV 152 (lane 2) and HCV 173 (lane 3) showing migration positions of HCV genotypes 2a (190 bp) and 2b (337 bp). Two DNA bands with sizes corresponding to genotypes 2a and 2b observed in sample from donor labelled HCV 152 indicated this donor had dual genotype infection. (c) Semi-nested PCR amplification products (429 bp) from positive samples (HCV 152 and 173) purified for cloning and sequencing. (d) Five products [(HCV 173–1, 173–2, HCV 152–4, 152–5 and 152–8 of Eco RI digested rDNA (4.35 kb) clones purified from E. coli cultures. Positive clones showed two bands of sizes 3.9 kb and 429 bp corresponding to the vector and insert cDNA respectively