Difference between revisions of "TaqMan SNP Assay"
From Bridges Lab Protocols
| Line 15: | Line 15: | ||
== Protocol == | == Protocol == | ||
# Dilute gDNA samples to 1-5 ng/ul. | # Dilute gDNA samples to 1-5 ng/ul. | ||
| − | # Place gDNA, TaqMan® Genotyping Master Mix, TaqMan® Genotyping Assay on ice. | + | # Place gDNA, TaqMan® Genotyping Master Mix (40X), TaqMan® Genotyping Assay (2X) on ice. |
# Make up Reaction Master mix (see table below). | # Make up Reaction Master mix (see table below). | ||
| − | + | {| class="wikitable" | |
| + | |- | ||
| + | ! Component | ||
| + | ! 1X volume | ||
| + | ! 96X Volume (+ 10% excess) | ||
| + | |- | ||
| + | |TaqMan® Genotyping Master Mix (2X) | ||
| + | |TaqMan® Genotyping Assay (40X) | ||
| + | |- | ||
| + | |5.00 ul | ||
| + | |0.25 ul | ||
| + | |- | ||
| + | |528.0 ul | ||
| + | |26.4 ul | ||
Revision as of 16:14, 16 November 2015
Materials
- 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)
Protocol
- Dilute gDNA samples to 1-5 ng/ul.
- Place gDNA, TaqMan® Genotyping Master Mix (40X), TaqMan® Genotyping Assay (2X) on ice.
- Make up Reaction Master mix (see table below).
| Component | 1X volume | 96X Volume (+ 10% excess) |
|---|---|---|
| TaqMan® Genotyping Master Mix (2X) | TaqMan® Genotyping Assay (40X) | |
| 5.00 ul | 0.25 ul | |
| 528.0 ul | 26.4 ul |
