Heteroduplex analysis

Heteroduplexes and homoduplexes formed after amplification and re-annealing of wild type and mutant alleles

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. ^ a b J. Wallace, Andrew (2002). "SSCP/Heteroduplex Analysis". In D. M. Theophilus, Bimal; Rapley, Ralph (eds.). PCR Mutation Detection Protocols. Totowa, New Jersey: Humana Press, Totowa, New Jersey. pp. 151 - 164. ISBN 0896036170.
  2. ^ Glavač, Damjan; Dean, Michael (1996), Pfeifer, Gerd P. (ed.), "Heteroduplex Analysis", 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{{citation}}: CS1 maint: work parameter with ISBN (link)
  3. ^ El-Yazbi, Amira F.; Wong, Alysha; Loppnow, Glen R. (2017). "A luminescent probe of mismatched DNA hybridization: Location and number of mismatches". Analytica Chimica Acta. 994: 92–99. Bibcode:2017AcAC..994...92E. doi:10.1016/j.aca.2017.09.036. PMID 29126473.
  4. ^ 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. ^ a b 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 ed.), Academic Press, pp. 45–58, doi:10.1016/b978-0-12-374537-8.00004-3, ISBN 978-0-12-374537-8, retrieved 2019-12-02{{citation}}: CS1 maint: work parameter with ISBN (link)
  6. ^ a b 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". Journal of Biological Chemistry. 267 (7): 4911–4915. doi:10.1016/S0021-9258(18)42917-1. ISSN 0021-9258. PMID 1537869.

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