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SNPWaveTM: a flexible multiplexed SNP genotyping technology
Authors:Michiel J T van Eijk  José L N Broekhof  Hein J A van der Poel  René C J Hogers  Harrie Schneiders  Judith Kamerbeek  Esther Verstege  Joris W van Aart  Henk Geerlings  Jaap B Buntjer  A Jan van Oeveren  and Pieter Vos
Abstract:Scalable multiplexed amplification technologies are needed for cost-effective large-scale genotyping of genetic markers such as single nucleotide polymorphisms (SNPs). We present SNPWaveTM, a novel SNP genotyping technology to detect various subsets of sequences in a flexible fashion in a fixed detection format. SNPWave is based on highly multiplexed ligation, followed by amplification of up to 20 ligated probes in a single PCR. Depending on the multiplexing level of the ligation reaction, the latter employs selective amplification using the amplified fragment length polymorphism (AFLP®) technology. Detection of SNPWave reaction products is based on size separation on a sequencing instrument with multiple fluorescence labels and short run times. The SNPWave technique is illustrated by a 100-plex genotyping assay for Arabidopsis, a 40-plex assay for tomato and a 10-plex assay for Caenorhabditis elegans, detected on the MegaBACE 1000 capillary sequencer.
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