Assessment of high resolution melting analysis as a potential SNP genotyping technique in forensic casework

Bhavik MEHTA, Runa Daniel, Roland van Oorschot, Dennis MCNEVIN

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

High resolution melting (HRM) analysis is a simple, cost effective, closed tube SNP genotyping technique with high throughput potential. The effectiveness of HRM for forensic SNP genotyping was assessed with five commercially available HRM kits evaluated on the ViiA™ 7 Real Time PCR instrument. Four kits performed satisfactorily against forensically relevant criteria. One was further assessed to determine the sensitivity, reproducibility, and accuracy of HRM SNP genotyping. The manufacturer's protocol using 0.5 ng input DNA and 45 PCR cycles produced accurate and reproducible results for 17 of the 19 SNPs examined. Problematic SNPs had GC rich flanking regions which introduced additional melting domains into the melting curve (rs1800407) or included homozygotes that were difficult to distinguish reliably (rs16891982; a G to C SNP). A proof of concept multiplexing experiment revealed that multiplexing a small number of SNPs may be possible after further investigation. HRM enables genotyping of a number of SNPs in a large number of samples without extensive optimization. However, it requires more genomic DNA as template in comparison to SNaPshot®. Furthermore, suitably modifying pre-existing forensic intelligence SNP panels for HRM analysis may pose difficulties due to the properties of some SNPs
Original languageEnglish
Pages (from-to)3036-3043
Number of pages8
JournalElectrophoresis
Volume35
Issue number21-22
DOIs
Publication statusPublished - 2014

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Genotyping Techniques
Freezing
Single Nucleotide Polymorphism
Melting
Multiplexing
GC Rich Sequence
DNA
Homozygote
Intelligence
Throughput
Real-Time Polymerase Chain Reaction

Cite this

MEHTA, Bhavik ; Daniel, Runa ; van Oorschot, Roland ; MCNEVIN, Dennis. / Assessment of high resolution melting analysis as a potential SNP genotyping technique in forensic casework. In: Electrophoresis. 2014 ; Vol. 35, No. 21-22. pp. 3036-3043.
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Assessment of high resolution melting analysis as a potential SNP genotyping technique in forensic casework. / MEHTA, Bhavik; Daniel, Runa; van Oorschot, Roland; MCNEVIN, Dennis.

In: Electrophoresis, Vol. 35, No. 21-22, 2014, p. 3036-3043.

Research output: Contribution to journalArticle

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