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*TaqMan® Genotyping Assay (supplied at 20X, 40X or 80X) (Applied Biosystems).
*TaqMan TaqMan® Genotyping Master Mix (supplied at 2X)(Applied Biosystems).
*StepOnePlus Real-Time PCR System (Applied Biosystems).
*PC/Laptop with StepOnePlus Software installed.
*Optical Plate Covers.
*Nuclease-Free Water
*Sample Genomic DNA (extracted as described in [[Blood DNA Extraction]])
*Postive control(s) (Minor allele) (''optional, but recommended'')
== Protocol ==
# Dilute gDNA samples to 20ng/ul approx.
# Place gDNA, TaqMan® Genotyping Master Mix (40X), TaqMan® Genotyping Assay (2X) on ice.
# Make up Reaction Master mix (TaqMan® Genotyping Master Mix (40X), TaqMan® Genotyping Assay (2X): see table below).
# Add Reaction Mix (5.25 ul) added to each well (Table 1.)
# Diluted gDNA samples each added to a single well (Table 1.)
'''Table 1. Full 96 well plate'''
{| class="wikitable"
|-
! Component
! 1X volume
! 96X Volume (+ 10% excess)
|-
|TaqMan® Genotyping Master Mix (2X)
|5.00 ul
|528.0 ul
|-
|TaqMan® Genotyping Assay (40X)
|0.25 ul
|26.4 ul
|-
|gDNA Sample (20ng/ul)
|4.50 ul
|501.6 ul
|-
!
!10 ul
!1056 ul
|}