Abstract
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.
Keywords
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References
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Article info
Publication history
Published online: September 03, 2008
Accepted:
June 30,
2008
Received in revised form:
June 19,
2008
Received:
April 11,
2008
Identification
Copyright
© 2008 Elsevier Ireland Ltd. Published by Elsevier Inc. All rights reserved.
