Abstract
We report the development of an effective system for analyzing X chromosome-linked mini short tandem repeat loci with reduced-size amplicons (less than 220 bp), useful for analyzing highly degraded DNA samples. To generate smaller amplicons, we redesigned primers for eight X-linked microsatellites (DXS7132, DXS10079, DXS10074, DXS10075, DXS6801, DXS6809, DXS6789, and DXS6799) and established efficient conditions for a multiplex PCR system (miniX). The validation tests confirmed that it has good sensitivity, requiring as little as 20 pg of DNA, and performs well with DNA from paraffin-embedded tissues, thus showing potential for improved analysis and identification of highly degraded and/or very limited DNA samples. Consequently, this system may help to solve complex forensic cases, particularly when autosomal markers convey insufficient information.
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References
Carracedo A, Beckmann A, Bengs A et al (2001) Results of a collaborative study of the EDNAP group regarding the reproducibility and robustness of the Y chromosome STRs DYS 19, DYS 389 I and II, DYS 390 and DYS 393 in a PCR pentaplex format. Forensic Sci Int 119:28–41
Gusmão L, Alves C, Gomes I, Sánchez-Diz P (2012) Capillary electrophoresis of an X-chromosome STR decaplex for kinship deficiency cases. Methods Mol Biol 830:57–71
Szibor R, Krawczak M, Hering S, Edelmann J, Kuhlisch E, Krause D (2003) Use of X-linked markers for forensic purposes. Int J Legal Med 117:67–74
Szibor R (2007) X-chromosomal markers: past, present and future. Forensic Sci Int Genet 1:93–99
Edelmann J, Hering S, Augustin C, Kalis S, Szibor R (2010) Validation of six closely linked STRs located in the X centromero region. Int J Legal Med 124:83–87
Rodig H, Kloep F, Weiaßach L, Augustin C, Edelmann J, Hering S, Szibor R, Götz F, Brabetz W (2010) Evaluation of seven X-chromosomal short tandem repeat loci located within the Xq26 region. Forensic Sci Int Genet 4:194–199
Szibor R, Hering S, Kuhlisch E, Plate I, Demberger S, Krawczak M, Edelmann J (2005) Haplotyping of STR cluster DXS6801-DXS6809-DXS6789 on Xq21 provides a powerful tool for kinship testing. Int J Legal Med 119:363–369
Hering S, Augustin C, Edelmann J, Heidel M, Dreβler J, Rodig H, Kuhlisch E, Szibor R (2006) DXS10079, DXS10074 and DXS10075 are STRs located within a 280-kb region of Xq12 and provide stable haplotypes useful for complex kinship cases. Int J Legal Med 120:337–345
Butler JM, Shen Y, McCord BR (2003) The development of reduced size STR amplicons as tool for analysis of degraded DNA. J Forensic Sci 48:1054–1064
Coble MD, Butler JM (2005) Characterization of new miniSTR loci to aid analysis of degraded DNA. J Forensic Sci 50:43–53
Castañeda M, Mijares V, Riancho JA, Zarrabeitia MT (2012) Haplotypic blocks of X-linked STRs for forensic cases: study of recombination and mutation rates. J Forensic Sci 57(1):192–195
Hering S, Rodig H, Augustin C, Edelmann J, Heidel M, Dressler J, Szibor R (2007) DXS10079, DXS10074 and DXS10075: new alleles and SNP occurrence. EXCLI J 6:177–182
Brownstein MJ, Carpten JR, Smith JR (1996) Modulation of non-templated nucleotide addition by Taq DNA polymerase: primer modifications that facilitate genotyping. Biotechniques 20:1004–1010
Gusmão L, Sánchez-Diz P, Alves C, Gomes I, Zarrabeitia MT et al (2009) A GEP-ISFG collaborative study on the optimization of an X-STR decaplex: data on 15 Iberian and Latin American populations. Int J Legal Med 123(3):227–234
Asamura H, Sakai H, Kobayashi K, Ota M, Fukushima H (2006) MiniX-STR multiplex system population study in Japan and application to degraded DNA analysis. Int J Legal Med 120:174–181
Diegoli TM, Coble MD (2011) Development and characterization of two miniX chromosomal short tandem repeat multiplexes. Forensic Sci Int Genet 5(5):415–421
Walsh SP, Fildes NJ, Reynolds R (1996) Sequence analysis and characteristics of stutter products at the tetranucleotide repeat locus vWA. Nucleic Acids Res 24(14):2807–2812
Odriozola A, Aznar JM, Celorrio D, Bravo ML, Builes JJ, Val-Bernal JF, de Pancorbo MM (2011) Development and validation of I-DNA1: a 15-loci multiplex system for identity testing. Int J Legal Med 125:685–694
Odriozola A, Aznar JM, Celorrio D, Bravo ML, Builes JJ, De Pancorbo MM (2012) Development and validation for identity testing of I-DNADuo, a combination of I-DNA1 and a new multiplex system, I-DNA2. Int J Legal Med 126:167–172
Schelenk J, Seidl S, Braunschweiger G, Betz P, Lederer T (2004) Development of a 13-locus PCR multiplex system for paternity testing. Int J Legal Med 118:55–61
Mulero JJ, Chang CW, Lagacé RE, Wang DY, Bas JL, McMahon TP, Hennessy LK (2008) Development and validation of the AmpFlSTR® MiniFiler™ PCR amplification kit: a MiniSTR multiplex for the analysis of degraded and/or PCR inhibited DNA. J Forensic Sci 53(4):838–852
Wang DY, Chang CW, Lagacé RE, Oldroyd NJ, Hennessy LK (2011) Development and validation of the AmpFlSTR® Identifiler® Direct PCR Amplification Kit: a multiplex assay for the direct amplification of single-source samples. J Forensic Sci 56(4):835–845
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This study was supported in part by a grant from IFIMAV (API 11/26).
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Castañeda, M., Odriozola, A., Gómez, J. et al. Development and validation of a multiplex reaction analyzing eight miniSTRs of the X chromosome for identity and kinship testing with degraded DNA. Int J Legal Med 127, 735–739 (2013). https://doi.org/10.1007/s00414-012-0795-2
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DOI: https://doi.org/10.1007/s00414-012-0795-2