Fluidigm Juno - Prepare the Sample MIX

Genotyping with Juno Getting Started Guide
Chapter 2: Genotyping with the Juno 96.96 Genotyping IFC Using TaqMan Assays
Prepare Assay and Sample Mixes
15
Prepare the Sample Mix
1 In a DNA-free hood, in a new 1.5-mL microcentrifuge tube labeled “Sample Pre-Mix,”
combine the Juno GT Preamp Master Mix and the primer pool for preamplification to
prepare the sample pre-mix. (See Table 3.)
2 Label a new 96-well plate “SAMPLE PLATE.” Pipet 2.25 μL of the sample pre-mix
into each well of the plate. Skip wells that are for no template controls. Do not add
sample pre-mix to no template control wells.
IMPORTANT Prepare at least one no template control.
3 In a DNA sample hood, pipet 2.75 μL of genomic DNA into the appropriate wells of
the sample plate.
4 In a DNA sample hood, pipet 5.00 μL of Dilution Reagent into each no template
control well.
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.
Table 3. Sample mix
IMPORTANT Do not go past the first stop on the pipette. Doing so may introduce air
bubbles into the inlets. To avoid bubbles, pipet 4.0 μL into each inlet from the 5.0 μL
overage volume.
Component Volume per
Inlet (μL)
Volume per
Inlet with
Overage
(μL)
Sample Mix
for IFC with
Overage*
(μL)
* 120 reactions for ease of pipetting
SAMPLE PRE-MIX
Juno GT Preamp Master Mix
0.8 1.00 120.0
Primer pool for preamplification†
† See “Prepare the Primer Pool for Preamplification” on page 13.
1.0 1.25 150.0
Genomic DNA
2.2 2.75 —
Total
4.0 5.0 270.0