Abstract
We describe a method for the simultaneous typing of Y-chromosome single nucleotide polymorphism (SNP) markers by means of multiplex polymerase chain reaction (PCR) strategies that allow the detection of 35 Y chromosome SNPs on 25 amplicons from 100 to 200 pg of chromosomal deoxyribonucleic acid (DNA). Multiplex PCR amplification of the DNA was performed with slight modifications of standard PCR conditions. Single-base extension (SBE) was performed using the SNaPshot kit containing fluorescently labeled ddNTPs. The extended primers were detected on an ABI 3100 sequencer. The most important factors for the creation of larger SNP typing PCR multiplexes include careful selection of primers for the primary amplification and the SBE reaction, use of DNA primers with homogenous composition, and balancing the primer concentrations for both the amplification and the SBE reactions.
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© 2005 Humana Press Inc., Totowa, NJ
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Sanchez, J.J., Børsting, C., Morling, N. (2005). Typing of Y Chromosome SNPs With Multiplex PCR Methods. In: Carracedo, A. (eds) Forensic DNA Typing Protocols. Methods in Molecular Biology, vol 297. Humana Press. https://doi.org/10.1385/1-59259-867-6:209
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DOI: https://doi.org/10.1385/1-59259-867-6:209
Publisher Name: Humana Press
Print ISBN: 978-1-58829-264-3
Online ISBN: 978-1-59259-867-0
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