GenPlex (Applied Biosystems) is a new SNP genotyping system based on an initial PCR amplification followed by an oligo ligation assay (OLA). The OLA consists of the hybridization of allele and locus specific oligonucleotides (ASOs and LSOs) to PCR products and posterior ligation of ASOs and LSOs. The ligation products are immobilized to microtitre plates and reporter oligonucleotides (ZipChute® probes) are hybridized to the ligation products. ZipChute® probes are subsequently eluted and detected using capillary electrophoresis. Applied Biosystems developed the GenPlex SNP genotyping system with amelogenin and 48 of the 52 SNPs used in the 52 SNP-plex assay developed by the SNPforID consortium. The system requires equipment that is usually found in forensic genetic laboratories. The use of a robot for performance of the pipetting steps is highly recommendable.
A total of 286 individuals from Denmark, Somalia and Greenland were investigated with GenPlex using a Biomek® 3000 (Beckman Coulter) robot. The results were compared to results obtained with an ISO 17025 accredited SNP typing assay based on single base extension (SBE). With the GenPlex SNP genotyping system, full SNP profiles were obtained in 97.6% of the investigations. Perfect concordance was obtained in duplicate investigations and the SNP genotypes obtained with the GenPlex system were concordant with those of the accredited SBE based SNP typing system except for one result in rs901398 in one of 286 individuals most likely due to a mutation 6 bp downstream of the SNP. Reproducible SNP genotypes were obtained from as little as 250 pg of DNA.
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The SNPforID consortium: http://www.snpforid.org.
- A multiplex assay with 52 single nucleotide polymorphisms for human identification.Electrophoresis. 2006; 27 (9): 1713-1724
- Performance of the SNPforID 52 SNP-plex assay in paternity testing.Forensic Sci. Int. 2008; 2: 292-300
- DNA polymerase-catalyzed addition of nontemplated extra nucleotides to the 3’ end of a DNA fragment.DNA Cell. Biol. 1993; 12: 763-770
- Substrate nucleotide-determined non-templated addition of adenine by Taq DNA polymerase: implications for PCR-based genotyping and cloning.Biotechniques. 1996; 21: 700-709
- Typing of multiple SNPs Using ribonuclease cleavage of DNA/RNA chimeric single base extension primers and detection by MALDI-TOF mass spectrometry.Anal. Chem. 2005; 77: 5229-5235
- SNP typing on the NanoChip electronic microarray.Methods Mol. Biol. 2005; 297: 155-168
- A sensitive issue: pyrosequencing as a valuable forensic SNP typing platform.Int. Congress Ser. 2006; 1288: 52-54
- SNP genotyping with single base extension-tag microarrays.Int. Congress Ser. 2004; 1261: 331-333
- SNPs in forensic genetics: a review of SNP typing methodologies.Forensic Sci. Int. 2005; 154: 181-191
AB SNPlex™ chemistry guide pdf: http://docs.appliedbiosystems.com/pebiodocs/04360856.pdf.
- Evaluation of the Genplex SNP typing system and a 49-plex forensic marker panel.Forensic Sci. Int. Genet. 2007; 1: 180-185
- Biomek®-3000 and GenPlex SNP genotyping in forensic genetics.JALA (J. Assoc. Lab. Autom.). 2008; 13: 297-303
Published online: September 03, 2008
Accepted: June 30, 2008
Received in revised form: June 19, 2008
Received: April 11, 2008
© 2008 Elsevier Ireland Ltd. Published by Elsevier Inc. All rights reserved.