Genotyping with Juno Getting Started Guide
13
Chapter 2: Genotyping with the
Juno 96.96 Genotyping IFC Using
TaqMan Assays
Prepare Assay and Sample Mixes
Prepare the Primer Pool for Preamplification
1 If necessary, adjust the concentration of TaqMan genotyping assays with DNase-free
water to 18 μM (20X).
2 In a new, labeled 1.5-mL microcentrifuge tube, combine 2 μL of each 20X TaqMan
genotyping assay up to a total of 96 assays. The total volume of assays is 2Y in
Table 1, where Y is the number of assays used. Each assay is at a final concentration
of 0.2X in the primer pool.
3 Add Dilution Reagent to the 20X TaqMan assays:
Table 1. Prepare the primer pool for preamplification
The final concentration of each primer in the preamplification reaction is 45 nM.
NOTE The volume can be adjusted proportionally based on the number of samples
to be amplified.
Component Volume (μL) Final Concentration
20X TaqMan genotyping assays,
18 μM*
* See step 1.
2Y (up to 96 assays) 180 nM (0.2X)
Dilution Reagent 200 – 2Y —
Total 200.0 —
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