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Differentiating between monozygotic twins through DNA methylation-specific high-resolution melt curve analysis

Stewart, Leander; Evans, Neil; Bexon, Kimberley J.; Van Der Meer, Dieudonne J.; Williams, Graham A.


Leander Stewart

Neil Evans

Kimberley J. Bexon

Dieudonne J. Van Der Meer


Although short tandem repeat profiling is extremely powerful in identifying individuals from crime scene stains, it is unable to differentiate between monozygotic (MZ) twins. Efforts to address this include mutation analysis through whole genome sequencing and through DNA methylation studies. Methylation of DNA is affected by environmental factors; thus, as MZ twins age, their DNA methylation patterns change. This can be characterized by bisulfite treatment followed by pyrosequencing. However, this can be time-consuming and expensive; thus, it is unlikely to be widely used by investigators. If the sequences are different, then in theory the melting temperature should be different. Thus, the aim of this study was to assess whether high-resolution melt curve analysis can be used to differentiate between MZ twins. Five sets of MZ twins provided buccal swabs that underwent extraction, quantification, bisulfite treatment, polymerase chain reaction amplification and high-resolution melting curve analysis targeting two markers, Alu-E2F3 and Alu-SP. Significant differences were observed between all MZ twins targeting Alu-E2F3 and in four of five MZ twins targeting Alu-SP (P < 0.05). Thus, it has been demonstrated that bisulfite treatment followed by high-resolution melting curve analysis could be used to differentiate between MZ twins.


Stewart, L., Evans, N., Bexon, K. J., Van Der Meer, D. J., & Williams, G. A. (2015). Differentiating between monozygotic twins through DNA methylation-specific high-resolution melt curve analysis. Analytical Biochemistry, 476, 36-39.

Journal Article Type Article
Acceptance Date Feb 2, 2015
Online Publication Date Feb 10, 2015
Publication Date May 1, 2015
Deposit Date May 16, 2022
Journal Analytical Biochemistry
Print ISSN 0003-2697
Electronic ISSN 1096-0309
Publisher Elsevier
Peer Reviewed Peer Reviewed
Volume 476
Pages 36-39
Keywords Forensic genetics; DNA methylation; Monozygotic twins; High-resolution melt curve analysis
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