Fluidigm Juno - Prepare 2 X Taqman Assays for Genotyping; Prepare the Assay MIX; Obtain the Minimum Required Genomic DNA

Genotyping with Juno Getting Started Guide
Chapter 2: Genotyping with the Juno 96.96 Genotyping IFC Using TaqMan Assays
Prepare Assay and Sample Mixes
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Prepare 2X TaqMan Assays for Genotyping
1 If necessary, adjust the concentration of TaqMan genotyping assays with DNase-free
water to 18 μM (20X).
2 In a new 96-well plate, dilute the 20X TaqMan genotyping assays in Dilution
Reagent or DNase-free water to a final concentration of 2X for each assay:
Prepare the Assay Mix
1 Label a new 96-well plate, “TAQMAN ASSAY PLATE.” In a DNA-free hood, pipet
2.5 μL of Probe GT Master Mix into each well. (See Table 2.)
2 In a DNA-free hood, pipet 2.5 μL of 2X TaqMan assays into a well of the TaqMan
assay plate for each assay. (See “Prepare 2X TaqMan Assays for Genotyping”.)
3 In unused assay inlets, combine 2.5 μL of Probe GT Master Mix with 2.5 μL DNase-
free water.
4 Seal the plate with MicroAmp Clear Adhesive Film, vortex it for 5 seconds, then
centrifuge it at 1,000 x g for 1 minute.
Table 2. Assay mix
Obtain the Minimum Required Genomic DNA
For high-quality human samples, the minimum DNA required is 2.5 ng/μL in 2.75 μL.
Larger genomes require higher concentrations of genomic DNA.
Component Volume (μL) Final Concentration
20X TaqMan genotyping assays 1.0 2X
Dilution Reagent or
DNase-free water
9.0 —
Total 10.0 —
Component Volume (μL)
Probe GT Master Mix 2.5
2X TaqMan assays*
* See “Prepare 2X TaqMan Assays for Genotyping”.
2.5
Total 5.0