DefinePK

Abstract

Objective: The current study was aimed at analyzing the utility of intragenic polymorphic restriction sites (HindIII and BclI) in screening of haemophilia A patients and their immediate family members.
Methodology: Thirteen families having one or more affected child were recruited from Pakistan Institute of Medical Sciences. For Genomic DNA extraction, phenol-chloroform method was used. Genomic regions containing the RFLP site were amplified by using specific primers designer from flanking regions. The PCR products were digested using HindIII and BclI and the products were resolved on 6% Polyacrylamide gel electrophoresis and visualized by silver staining technique.
Results: A total of 100 X-chromosomes were investigated in this study (36 males, 32 females). The minor allele frequency for Intron 18/BclI T>A was observed to be 0.68 while for Intron 19/HindIII C>T it was observed to be 0.55 in our study population. The Intron 18/BclI T>A marker was informative in four of the families while Intron 19/HindIII C>T was informative for 8 families. The PIC was calculated to be 0.34 for BclI marker and 0.37 for HindIII .
Conclusion: The rate of heterozygosity and the PIC values obtained for the two markers suggests that HindIII marker is more informative as compared to BclI in our study population. The two markers were decisive in haplotyping 8 (66.6%) families.