Application of the Amplifluor SNPs genotyping system using a BMG LABTECH microplate reader
SNPs (Single nucleotide polymorphisms) are positions in genomic DNA with a single nucleotide difference between individuals of a species.
In this application note we describe a homogeneous fluorescence assay based on PCR. The method utilizes two fluorescently labeled oligonucleotides that consist of a specific 3' primer sequence region and a 5' hairpin region that is labeled with either of two fluorophores (FAM or JOE). The presence of the 3' primer sequence enables these fluorescent oligo-nucleotides to work with target specific unlabeled primers designed to the researcher's specific target sequence. Thus, the method can be used for any variation of bases.
High S/B values indicate that the BMG LABTECH microplate reader is an excellent instrument for running this assay. It is easily capable of measuring the output from the Amplifluor SNP Genotyping assay kit (Millipore) with high precision and with high throughput.