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HindIII-based restriction fragment length polymorphism in hemophilic and nonhemophilic patients

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., rest...

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Autores principales: Dubey, Alok Kumar, Hussain, Nuzhat, Mittal, Neha
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Medknow Publications Pvt Ltd 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3217284/
https://www.ncbi.nlm.nih.gov/pubmed/22096332
http://dx.doi.org/10.4103/0976-9668.71669
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author Dubey, Alok Kumar
Hussain, Nuzhat
Mittal, Neha
author_facet Dubey, Alok Kumar
Hussain, Nuzhat
Mittal, Neha
author_sort Dubey, Alok Kumar
collection PubMed
description 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%.
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spelling pubmed-32172842011-11-17 HindIII-based restriction fragment length polymorphism in hemophilic and nonhemophilic patients Dubey, Alok Kumar Hussain, Nuzhat Mittal, Neha J Nat Sci Biol Med Medicine 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%. Medknow Publications Pvt Ltd 2010 /pmc/articles/PMC3217284/ /pubmed/22096332 http://dx.doi.org/10.4103/0976-9668.71669 Text en © Journal of Natural Science, Biology and Medicine http://creativecommons.org/licenses/by-nc-sa/3.0 This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Medicine
Dubey, Alok Kumar
Hussain, Nuzhat
Mittal, Neha
HindIII-based restriction fragment length polymorphism in hemophilic and nonhemophilic patients
title HindIII-based restriction fragment length polymorphism in hemophilic and nonhemophilic patients
title_full HindIII-based restriction fragment length polymorphism in hemophilic and nonhemophilic patients
title_fullStr HindIII-based restriction fragment length polymorphism in hemophilic and nonhemophilic patients
title_full_unstemmed HindIII-based restriction fragment length polymorphism in hemophilic and nonhemophilic patients
title_short HindIII-based restriction fragment length polymorphism in hemophilic and nonhemophilic patients
title_sort hindiii-based restriction fragment length polymorphism in hemophilic and nonhemophilic patients
topic Medicine
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3217284/
https://www.ncbi.nlm.nih.gov/pubmed/22096332
http://dx.doi.org/10.4103/0976-9668.71669
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