Genotyping with Juno Getting Started Guide
Chapter 2: Genotyping with the Juno 96.96 Genotyping IFC Using TaqMan Assays
Load and Run the IFC on Juno
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5 Load an entire syringe of Juno 96.96 GT Control Line Fluid into a reservoir and a
second syringe into the second reservoir. (See long pink rectangles on the right side
of the pipetting map.)
IMPORTANT Carefully dispense control line fluid into the reservoirs. If control line
fluid comes into contact with the sample inlets, use a new IFC.
6 Pipet 15 μL of Juno GT Flux Fluid into each of the six ports. (See purple circles on the
pipetting map.)
7 Unseal the TaqMan assay plate and pipet 4.0 μL of each assay mix into an assay
inlet. (See black circles on the pipetting map and “Prepare the Assay Mix” on
page 14.)
8 Unseal the sample plate and pipet 4.0 μL of each sample mix into a sample inlet.
(See green circles on the pipetting map and “Prepare the Sample Mix” on page 15.)
IMPORTANT Do not go past the first stop on the pipette. Doing so may introduce air
bubbles into the inlets. Pipet 4.0 μL from the 5.0 μL overage volume to ensure that
no air bubbles enter the inlet.
9 Pull the sticker front tab down and away from the IFC to gently peel off the loading
map. Do not invert the IFC.
10 If necessary, remove any bubbles from an IFC inlet by removing the contents by
pipette and then carefully re-pipetting the contents into the inlet.
11 Ensure that the SX interface plate (silver label) is installed in the instrument. [See the
Juno System User Guide (PN 100-7070).]
12 Place the IFC into the Juno instrument, then start the run <60 minutes after pipetting
the reagents into the IFC.
13 On the Juno Scripts screen, tap the Probe GT tab, Juno 96.96 Fast, then Run. It
takes ~2.7 hours to complete.
The script contains these thermal cycling protocols:
Cycles Temperature Time
Hot start 95 ºC 2 min
14 95 ºC 15 sec
60 ºC 4 min
Cycles Temperature Time
Hot start 95 ºC 2 min
45 95 ºC 2 sec
60 ºC 20 sec
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