Fluidigm Juno - Prepare the Assay MIX; Obtain the Minimum Required Genomic DNA

Genotyping with Juno Getting Started Guide
Chapter 3: Genotyping with the Juno 96.96 Genotyping IFC Using SNP Type Assays
Prepare Assay and Sample Mixes
22
Prepare the Assay Mix
1 In a DNA-free hood, in a new 1.5-mL microcentrifuge tube labeled “Assay Pre-Mix,”
combine the Juno SNP Type GT Master Mix and 60X SNP Type Reagent to prepare
the assay pre-mix. (See Table 5.)
2 Label a new 96-well plate “SNP TYPE ASSAY PLATE.” In a DNA-free hood, pipet
2.5 μL of the assay pre-mix into each well.
3 Pipet 2.5 μL of 2X SNP Type assay into each well of the SNP Type assay plate.
4 In unused assay or no-assay control inlets, combine 2.5 μL of assay pre-mix with
2.5 μL of Dilution Reagent.
5 Seal the plate with MicroAmp Clear Adhesive Film, vortex it for 5 seconds, then
centrifuge it at 1,000 x g for 1 minute.
Obtain the Minimum Required Genomic DNA
For high-quality human samples, the minimum DNA required is 2.5 ng/μL. Larger
genomes require higher concentrations of genomic DNA.
Table 5 Assay mix
Component Volume per
Inlet (μL)
Volume per
Inlet with
Overage
(μL)
Assay Mix
for IFC with
Overage*
(μL)
* 120 reactions for ease of pipetting
ASSAY PRE-MIX
Juno SNP Type GT Master Mix
1.933 2.417 290.0
60X SNP Type Reagent
0.066 0.083 10.00
2X SNP Type assays†
† See “Prepare 2X SNP Type Assays” on page 21.
2.00 2.5 —
Total
4.00 5.00 300