SNPWave: a flexible multiplexed SNP genotyping technology.
Scalable multiplexed amplification technologies are needed for cost-effective large-scale genotyping of genetic markers such as single nucleotide polymorphisms (SNPs). We present SNPWave, 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.
Keygene NV, Agro Business Park 90, PO Box 216, 6700 AE Wageningen, The Netherlands. michiel.van-eijk@keygene.com
Michiel J T
MJ
van Eijk
José L N
JL
Broekhof
Hein J A
HJ
van der Poel
René C J
RC
Hogers
Harrie
H
Schneiders
Judith
J
Kamerbeek
Esther
E
Verstege
Joris W
JW
van Aart
Henk
H
Geerlings
Jaap B
JB
Buntjer
A Jan
AJ
van Oeveren
Pieter
P
Vos
D000483:Alleles; D000818:Animals; D017360:Arabidopsis / Q000235:genetics*; D017173:Caenorhabditis elegans / Q000235:genetics*; D004247:DNA / Q000032:analysis / Q000235:genetics; D015342:DNA Probes / Q000235:genetics; D005838:Genotype; D018551:Solanum lycopersicum / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D015342:DNA Probes; D004247:DNA