The aim of the project was to detect the polymorphisms in the apo A1 gene, which might possibly be associated with the risk of atherosclerosis in claudicants. This aim was achieved by amplification of the three exons and four introns of the apo A1 gene and followed by sequencing of the amplicons. The annealing temperature estimated by gradient PCRS for the coding and non-coding regions was 60.5°C, 61.2°C, 62.5°C, 59.7°C, 58.1°C and 57.8°C for 5’flanking, exon2/intronB/exon3, exon 3, intron C, exon 4 and 3’flanking, respectively. Altogether, 138 PCRs were undertaken. It was not possible to detect any high confidence polymorphism in the present sample size which was further reduced due to failed sequencing reactions in some samples. 0nly 16 out of 60 samples were successful for sequencing reaction. None of them could be successfully sequenced with their both forward and reverse primers. Any true estimates of the allele and genotype frequencies of the mutations cannot be made from the observed small sample size. Also, depending on the sample size and the sequencing results, any inference as to how these mutations could be associated with artherosclerosis in claudicants could not be made.
|Date of Award||2008|