Biology:Heteroduplex analysis
Heteroduplex analysis (HDA) is a method in biochemistry used to detect point mutations in DNA (Deoxyribonucleic acid) since 1992.[1] Heteroduplexes are dsDNA molecules that have one or more mismatched pairs, on the other hand homoduplexes are dsDNA which are perfectly paired.[1][2] This method of analysis depend up on the fact that heteroduplexes shows reduced mobility relative to the homoduplex DNA.[3] heteroduplexes are formed between different DNA alleles.[4] In a mixture of wild-type and mutant amplified DNA, heteroduplexes are formed in mutant alleles and homoduplexes are formed in wild-type alleles.[5] There are two types of heteroduplexes based on type and extent of mutation in the DNA. Small deletions or insertion create bulge-type heteroduplexes which is stable and is verified by electron microscope.[6] Single base substitutions creates more unstable heteroduplexes called bubble-type heteroduplexes, because of low stability it is difficult to visualize in electron microscopy.[5] HDA is widely used for rapid screening of mutation of the 3 bp p.F508del deletion in the CFTR gene.[6]
References
- ↑ 1.0 1.1 J. Wallace, Andrew (2002). "SSCP/Heteroduplex Analysis". in D. M. Theophilus, Bimal. PCR Mutation Detection Protocols. Totowa, New Jersey: Humana Press, Totowa, New Jersey. pp. 151 - 164. ISBN 0896036170. https://archive.org/details/pcrmutationdetec00theo_0/page/151.
- ↑ Glavač, Damjan; Dean, Michael (1996), Pfeifer, Gerd P., ed., "Heteroduplex Analysis" (in en), Technologies for Detection of DNA Damage and Mutations (Springer US): pp. 241–251, doi:10.1007/978-1-4899-0301-3_18, ISBN 978-1-4899-0301-3
- ↑ F. El-Yazbi, Amira; Wong, Alysha; R. Loppnow, Glen (2017). "A luminescent probe of mismatched DNA hybridization: Location and number of mismatches". Analytica Chimica Acta 994: 92–99. doi:10.1002/0471142905.hg0703s33. PMID 18428333.
- ↑ White, Marga Belle; Carvalho, Magda; Derse, David; O'Brien, Stephen J.; Dean, Michael (1992-02-01). "Detecting single base substitutions as heteroduplex polymorphisms". Genomics 12 (2): 301–306. doi:10.1016/0888-7543(92)90377-5. ISSN 0888-7543. PMID 1740339.
- ↑ 5.0 5.1 Menounos, Panayiotis G.; Patrinos, George P. (2010-01-01), Patrinos, George P.; Ansorge, Wilhelm J., eds., "Chapter 4 - Mutation Detection by Single Strand Conformation Polymorphism and Heteroduplex Analysis", Molecular Diagnostics (Second Edition) (Academic Press): pp. 45–58, doi:10.1016/b978-0-12-374537-8.00004-3, ISBN 978-0-12-374537-8, http://www.sciencedirect.com/science/article/pii/B9780123745378000043, retrieved 2019-12-02
- ↑ 6.0 6.1 Wang, Y. H.; Barker, P.; Griffith, J. (1992-03-05). "Visualization of diagnostic heteroduplex DNAs from cystic fibrosis deletion heterozygotes provides an estimate of the kinking of DNA by bulged bases." (in en). Journal of Biological Chemistry 267 (7): 4911–4915. doi:10.1016/S0021-9258(18)42917-1. ISSN 0021-9258. PMID 1537869. http://www.jbc.org/content/267/7/4911.
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