TaqMan SNP Assay

• TaqMan® Genotyping Assay (supplied at 20X, 40X or 80X) (Applied Biosystems).
• TaqMan® Genotyping Master Mix (supplied at 2X)(Applied Biosystems).
• StepOnePlus Real-Time PCR System (Applied Biosystems).
• PC/Laptop with StepOnePlus Software installed.
• MicroAmp Fast 96-Well Plates.
• Optical Plate Covers.
• Nuclease-Free Water
• Sample Genomic DNA (extracted as described in Blood DNA Extraction)
• Postive control(s) (Minor allele) (optional, but recommended)

1. Dilute gDNA samples to 20ng/ul approx.
2. Place gDNA, TaqMan® Genotyping Master Mix (40X), TaqMan® Genotyping Assay (2X) on ice.
3. Make up Reaction Master mix (TaqMan® Genotyping Master Mix (40X), TaqMan® Genotyping Assay (2X): see table below).
4. Add Reaction Mix (5.25 ul) added to each well (Table 1.)
5. Diluted gDNA samples each added to a single well (Table 1.)

Table 1. Full 96 well plate