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 dsDNAmolecules 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
^ abJ. 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. ISBN0896036170.
^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, ISBN978-1-4899-0301-3
