One high-throughput method to determine
SNP genotypes is SNPWave
(Keygene N.V.), which uses multiplex
oligonucleotide ligation amplification
of allele-specific probes coupled
with AFLP®-primer selective amplification.
Basic principle of SNPWave Multiplex
allele-discrimination based on ligation
using probes with unique stuffer (green).
padlock ligation probes are constructed
that are specific to the SNP and flanking
sequences. Locus-specific probes hybridize
to denatured genomic DNA. Allele‐specificity
is determined by the SNP at the 5'
end of the padlock-probe. Probes that
contain a 5' nucleotide complementary
to the SNP will be ligated and amplified
by PCR in subsequent reactions. Alternatively,
Genotyping (Applied Biosystems)
has been used to investigate SNPs
in a diverse research, including breast
cancer and to genotype plants.
GeneMarker software genotyping software
is designed for fast, accurate and
efficient analysis of SNPlex
data from MegaBACE®,
capillary electrophoresis systems.