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Published in: Forensic Science, Medicine and Pathology 3/2017

01-09-2017 | Original Article

HRM and SNaPshot as alternative forensic SNP genotyping methods

Authors: Bhavik Mehta, Runa Daniel, Dennis McNevin

Published in: Forensic Science, Medicine and Pathology | Issue 3/2017

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Abstract

Single nucleotide polymorphisms (SNPs) have been widely used in forensics for prediction of identity, biogeographical ancestry (BGA) and externally visible characteristics (EVCs). Single base extension (SBE) assays, most notably SNaPshot® (Thermo Fisher Scientific), are commonly used for forensic SNP genotyping as they can be employed on standard instrumentation in forensic laboratories (e.g. capillary electrophoresis). High resolution melt (HRM) analysis is an alternative method and is a simple, fast, single tube assay for low throughput SNP typing. This study compares HRM and SNaPshot®. HRM produced reproducible and concordant genotypes at 500 pg, however, difficulties were encountered when genotyping SNPs with high GC content in flanking regions and differentiating variants of symmetrical SNPs. SNaPshot® was reproducible at 100 pg and is less dependent on SNP choice. HRM has a shorter processing time in comparison to SNaPshot®, avoids post PCR contamination risk and has potential as a screening tool for many forensic applications.
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Literature
1.
2.
go back to reference Kayser M, de Knijff P. Improving human forensics through advances in genetics, genomics and molecular biology. Nature Rev Genet. 2011;12(3):179–92.CrossRefPubMed Kayser M, de Knijff P. Improving human forensics through advances in genetics, genomics and molecular biology. Nature Rev Genet. 2011;12(3):179–92.CrossRefPubMed
3.
go back to reference Mehta B, Daniel R, Phillips C, McNevin D. Forensically relevant SNaPshot® assays for human DNA SNP analysis: a review. Int J Legal Med. 2017;131(1):21–37.CrossRefPubMed Mehta B, Daniel R, Phillips C, McNevin D. Forensically relevant SNaPshot® assays for human DNA SNP analysis: a review. Int J Legal Med. 2017;131(1):21–37.CrossRefPubMed
4.
go back to reference Sanchez JJ, Phillips C, Børsting C, Balogh K, Bogus M, Fondevila M, et al. A multiplex assay with 52 single nucleotide polymorphisms for human identification. Electrophoresis. 2006;27(9):1713–24.CrossRefPubMed Sanchez JJ, Phillips C, Børsting C, Balogh K, Bogus M, Fondevila M, et al. A multiplex assay with 52 single nucleotide polymorphisms for human identification. Electrophoresis. 2006;27(9):1713–24.CrossRefPubMed
5.
go back to reference Phillips C, Salas A, Sánchez JJ, Fondevila M, Gómez-Tato A, Álvarez-Dios J, et al. Inferring ancestral origin using a single multiplex assay of ancestry-informative marker SNPs. Forensic Sci Int Genet. 2007;1(3–4):273–80.CrossRefPubMed Phillips C, Salas A, Sánchez JJ, Fondevila M, Gómez-Tato A, Álvarez-Dios J, et al. Inferring ancestral origin using a single multiplex assay of ancestry-informative marker SNPs. Forensic Sci Int Genet. 2007;1(3–4):273–80.CrossRefPubMed
6.
go back to reference Walsh S, Liu F, Ballantyne KN, Oven MV, Lao O, Kayser M. IrisPlex: a sensitive DNA tool for accurate prediction of blue and brown eye colour in the absence of ancestry information. Forensic Sci Int Genet. 2011;5:170–80.CrossRefPubMed Walsh S, Liu F, Ballantyne KN, Oven MV, Lao O, Kayser M. IrisPlex: a sensitive DNA tool for accurate prediction of blue and brown eye colour in the absence of ancestry information. Forensic Sci Int Genet. 2011;5:170–80.CrossRefPubMed
7.
go back to reference Mehta B, Daniel R, McNevin D. High resolution melting (HRM) of forensically informative SNPs. Forensic Sci Int Genet Supp Ser. 2013;4(1):e376–7.CrossRef Mehta B, Daniel R, McNevin D. High resolution melting (HRM) of forensically informative SNPs. Forensic Sci Int Genet Supp Ser. 2013;4(1):e376–7.CrossRef
8.
go back to reference Venables SJ, Mehta B, Daniel R, Walsh SJ, van Oorschot RAH, McNevin D. Assessment of high resolution melting analysis as a potential SNP genotyping technique in forensic casework. Electrophoresis. 2014;35(21–22):3036–43.CrossRefPubMed Venables SJ, Mehta B, Daniel R, Walsh SJ, van Oorschot RAH, McNevin D. Assessment of high resolution melting analysis as a potential SNP genotyping technique in forensic casework. Electrophoresis. 2014;35(21–22):3036–43.CrossRefPubMed
9.
go back to reference Reed GH, Kent JO, Wittwer CT. High resolution DNA melting analysis for simple and efficient molecular diagnostics. Pharmacogenomics. 2007;8(6):597–608.CrossRefPubMed Reed GH, Kent JO, Wittwer CT. High resolution DNA melting analysis for simple and efficient molecular diagnostics. Pharmacogenomics. 2007;8(6):597–608.CrossRefPubMed
10.
go back to reference Gundry CN, Vandersteen JG, Reed GH, Pryor RJ, Chen J, Wittwer CT. Amplicon melting analysis with labeled primers: a closed-tube method for differentiating homozygotes and heterozygotes. Clin Chem. 2003;49(3):396–406.CrossRefPubMed Gundry CN, Vandersteen JG, Reed GH, Pryor RJ, Chen J, Wittwer CT. Amplicon melting analysis with labeled primers: a closed-tube method for differentiating homozygotes and heterozygotes. Clin Chem. 2003;49(3):396–406.CrossRefPubMed
11.
go back to reference Liew M, Pryor R, Palais R, Meadows C, Erali M, Lyon E, et al. Genotyping of single-nucleotide polymorphisms by high-resolution melting of small amplicons. Clin Chem. 2004;50(7):1156–64.CrossRefPubMed Liew M, Pryor R, Palais R, Meadows C, Erali M, Lyon E, et al. Genotyping of single-nucleotide polymorphisms by high-resolution melting of small amplicons. Clin Chem. 2004;50(7):1156–64.CrossRefPubMed
12.
go back to reference Walsh S, Lindenbergh A, Zuniga SB, Sijenb T. Knijff Pd, Kayser M, et al. developmental validation of the IrisPlex system: determination of blue and brown iris colour for forensic intelligence. Forensic Sci Int Genet. 2011;5(5):464–71.CrossRefPubMed Walsh S, Lindenbergh A, Zuniga SB, Sijenb T. Knijff Pd, Kayser M, et al. developmental validation of the IrisPlex system: determination of blue and brown iris colour for forensic intelligence. Forensic Sci Int Genet. 2011;5(5):464–71.CrossRefPubMed
13.
go back to reference AppliedBiosystems. ABI PRISM® SNaPshot™ Multiplex Kit. P/N 4323357 Rev. B ed: Thermo Fisher Scientific. 2010;1–42. AppliedBiosystems. ABI PRISM® SNaPshot™ Multiplex Kit. P/N 4323357 Rev. B ed: Thermo Fisher Scientific. 2010;1–42.
14.
go back to reference Walsh S, Chaitanya L, Clarisse L, Wirken L, Draus-Barini J, Kovatsi L, et al. Developmental validation of the HIrisPlex system: DNA-based eye and hair colour prediction for forensic and anthropological usage. Forensic Sci Int Genet. 2014;9:150–61.CrossRefPubMed Walsh S, Chaitanya L, Clarisse L, Wirken L, Draus-Barini J, Kovatsi L, et al. Developmental validation of the HIrisPlex system: DNA-based eye and hair colour prediction for forensic and anthropological usage. Forensic Sci Int Genet. 2014;9:150–61.CrossRefPubMed
15.
go back to reference Westen AA, Matai AS, Laros JF, Meiland HC, Jasper M, de Leeuw WJ, et al. Tri-allelic SNP markers enable analysis of mixed and degraded DNA samples. Forensic Sci Int Genet. 2009;3(4):233–41.CrossRefPubMed Westen AA, Matai AS, Laros JF, Meiland HC, Jasper M, de Leeuw WJ, et al. Tri-allelic SNP markers enable analysis of mixed and degraded DNA samples. Forensic Sci Int Genet. 2009;3(4):233–41.CrossRefPubMed
16.
go back to reference Swen JJ, Baak-Pablo RF, Guchelaar H-J, van der Straaten T. Alternative methods to a TaqMan assay to detect a tri-allelic single nucleotide polymorphism rs757210 in the HNF1β gene. Clin Chem Lab Med. 2012;50(2):279–84.CrossRef Swen JJ, Baak-Pablo RF, Guchelaar H-J, van der Straaten T. Alternative methods to a TaqMan assay to detect a tri-allelic single nucleotide polymorphism rs757210 in the HNF1β gene. Clin Chem Lab Med. 2012;50(2):279–84.CrossRef
17.
go back to reference Fondevila M, Phillips C, Santos C, Freire Aradas A, Vallone PM, Butler JM, et al. Revision of the SNPforID 34-plex forensic ancestry test: assay enhancements, standard reference sample genotypes and extended population studies. Forensic Sci Int Genet. 2013;7(1):63–74.CrossRefPubMed Fondevila M, Phillips C, Santos C, Freire Aradas A, Vallone PM, Butler JM, et al. Revision of the SNPforID 34-plex forensic ancestry test: assay enhancements, standard reference sample genotypes and extended population studies. Forensic Sci Int Genet. 2013;7(1):63–74.CrossRefPubMed
18.
go back to reference Seipp MT, Durtschi JD, Voelkerding KV, Wittwer CT. Multiplex amplicon genotyping by high-resolution melting. J Biomol Tech. 2009;20(3):160–4.PubMedPubMedCentral Seipp MT, Durtschi JD, Voelkerding KV, Wittwer CT. Multiplex amplicon genotyping by high-resolution melting. J Biomol Tech. 2009;20(3):160–4.PubMedPubMedCentral
19.
go back to reference Nicklas JA, Buel E. Improving the efficiency of DNA casework analysis through simple, effective, PCR-based screening methods. NIJ Grant#2005-DA-BX-K003. 2011;1–115. Nicklas JA, Buel E. Improving the efficiency of DNA casework analysis through simple, effective, PCR-based screening methods. NIJ Grant#2005-DA-BX-K003. 2011;1–115.
20.
go back to reference Derzelle S, Laroche S, Le Fleche P, Hauck Y, Thierry S, Vergnaud G, et al. Characterization of genetic diversity of bacillus anthracis in France by using high-resolution melting assays and multilocus variable-number tandem-repeat analysis. J Clin Microbiol. 2011;49(12):4286–92.CrossRefPubMedPubMedCentral Derzelle S, Laroche S, Le Fleche P, Hauck Y, Thierry S, Vergnaud G, et al. Characterization of genetic diversity of bacillus anthracis in France by using high-resolution melting assays and multilocus variable-number tandem-repeat analysis. J Clin Microbiol. 2011;49(12):4286–92.CrossRefPubMedPubMedCentral
21.
go back to reference Bosmali I, Ganopoulos I, Madesis P, Tsaftaris A. Microsatellite and DNA-barcode regions typing combined with high resolution melting (HRM) analysis for food forensic uses: a case study on lentils (Lens culinaris). Food Res Int. 2012;46(1):141–7.CrossRef Bosmali I, Ganopoulos I, Madesis P, Tsaftaris A. Microsatellite and DNA-barcode regions typing combined with high resolution melting (HRM) analysis for food forensic uses: a case study on lentils (Lens culinaris). Food Res Int. 2012;46(1):141–7.CrossRef
22.
go back to reference Ganopoulos I, Bazakos C, Madesis P, Kalaitzis P, Tsaftaris A. Barcode DNA high-resolution melting (bar-HRM) analysis as a novel close-tubed and accurate tool for olive oil forensic use. J Sci Food Agr. 2013;93(9):2281–6.CrossRef Ganopoulos I, Bazakos C, Madesis P, Kalaitzis P, Tsaftaris A. Barcode DNA high-resolution melting (bar-HRM) analysis as a novel close-tubed and accurate tool for olive oil forensic use. J Sci Food Agr. 2013;93(9):2281–6.CrossRef
23.
go back to reference Malewski T, Draber-Mońko A, Pomorski J, Łoś M, Bogdanowicz W. Identification of forensically important blowfly species (Diptera: Calliphoridae) by high-resolution melting PCR analysis. Int J Legal Med. 2010;124(4):277–85.CrossRefPubMed Malewski T, Draber-Mońko A, Pomorski J, Łoś M, Bogdanowicz W. Identification of forensically important blowfly species (Diptera: Calliphoridae) by high-resolution melting PCR analysis. Int J Legal Med. 2010;124(4):277–85.CrossRefPubMed
24.
go back to reference Sijen T. Molecular approaches for forensic cell type identification: on mRNA, miRNA, DNA methylation and microbial markers. Forensic Sci Int Genet. 2015;18:21–32.CrossRefPubMed Sijen T. Molecular approaches for forensic cell type identification: on mRNA, miRNA, DNA methylation and microbial markers. Forensic Sci Int Genet. 2015;18:21–32.CrossRefPubMed
Metadata
Title
HRM and SNaPshot as alternative forensic SNP genotyping methods
Authors
Bhavik Mehta
Runa Daniel
Dennis McNevin
Publication date
01-09-2017
Publisher
Springer US
Published in
Forensic Science, Medicine and Pathology / Issue 3/2017
Print ISSN: 1547-769X
Electronic ISSN: 1556-2891
DOI
https://doi.org/10.1007/s12024-017-9874-5

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