title: HindIII-based restriction fragment length polymorphism in hemophilic and nonhemophilic patients.
abstract: Hemophilia A is most common recessively inherited bleeding disorder, which affect one in five thousand male births throughout the world. In most of the hemophilic A patients, no common mutation is easily identifiable. This limitation has been overcome by the use of polymorphic DNA marker, i.e., restriction fragment length polymorphism (RFLP). This marker of polymorphism could only be detected by amplifying the polymorphic region and digestion the polymerase chain reaction (PCR) product with the restriction enzyme (PCR-RFLP), i.e., HindIII. The polymorphic region of HindIII is 608 bp in length and after the restriction digestion, different sizes of fragments, i.e., 427 and 181 bp were, respectively, obtained. However, in homozygous (+/+) condition three bands of 427, 100, and 81 bp were obtained and in the other negative allelic homozygous condition (-/-) two bands of 427 and 181 bp were obtained. Similarly fragments of different sizes, i.e., 427, 181, 100, and 81 bp were obtained in heterozygous conditions. Therefore, in this study, we have analyzed the factor VIII gene in the 17 different families using restriction enzyme HindIII-based RFLP molecular marker technique. Out of these, the observed heterozygosity for HindIII was found 47.5%, whereas, for positive allele it was 26%, and for negative allele the frequency was 74%.
authors.affiliation: Division of Cellulose and Paper, Forest Research Institute, P.O., IPE, Kaulagarh Road, Dehradun, Uttarakhand, India.
authors.forename: Alok Kumar
authors.initials: AK
authors.lastname: Dubey
authors.forename: Nuzhat
authors.initials: N
authors.lastname: Hussain
authors.forename: Neha
authors.initials: N
authors.lastname: Mittal
references.citation: Nat Genet. 1993 Nov;5(3):236-41
references.pmid: 8275087
references.citation: N Engl J Med. 1985 Oct 3;313(14):842-8
references.pmid: 2993888
references.citation: Br J Haematol. 1990 Sep;76(1):75-9
references.pmid: 1977469
references.citation: Am J Hum Genet. 1992 Jul;51(1):66-80
references.pmid: 1609806
references.citation: Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8307-11
references.pmid: 1924291
references.citation: Mol Pathol. 2002 Apr;55(2):127-44
references.pmid: 11950963
references.citation: Nature. 1984 Nov 22-28;312(5992):326-30
references.pmid: 6438525
references.citation: Proc Natl Acad Sci U S A. 1968 Apr;59(4):1300-6
references.pmid: 4870862
references.citation: Nucleic Acids Res. 1991 Sep 25;19(18):4821-33
references.pmid: 1923751
references.citation: Hum Genet. 1988 Apr;78(4):359-62
references.pmid: 2896159