Calibrate The Instrument; Prepare The Plate Or Array Card - Applied Biosystems ViiA 7 Real Quick Reference Manual

Fast 96-well sample block
• Fast 96-Well Spectral Calibration Plate with FAM
• Fast 96-Well Spectral Calibration Plate with VIC
• Fast 96-Well Spectral Calibration Plate with ROX
• Fast 96-Well Spectral Calibration Plate with SYBR
• Fast 96-Well Spectral Calibration Plate with TAMRA
• Fast 96-Well Spectral Calibration Plate with NED
• Fast 96-Well Region of Interest (ROI) and Background Plates (PN 4432426)
• Fast 96-Well Normalization Plates with FAM
(PN 4432432)
®
• TaqMan RNase P Fast 96-Well Instrument Verification Plate (PN 4351979)

Calibrate the instrument

1 Prepare the plate or
array card
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Applied Biosystems ViiA 7 Real-Time PCR System Quick Reference Guide
™
Dye (PN 4432389)
®
Dye (PN 4432396)
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Dye (PN 4432402)
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Green Dye (PN 4432408)
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Dye (PN 4432414)
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Dye (PN 4432420)
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/ROX and VIC
To prepare the calibration plate:
a.
Remove the plate from the freezer, then allow it to thaw to room temperature (approximately 5
minutes).
IMPORTANT!
Do not remove a plate from its packaging until you are ready to run it.
b.
Remove the plate from its packaging.
IMPORTANT! Do not discard the packaging. The plate can be used up to three times if it is
stored in its original packaging sleeve.
c.
Vortex the plate for 5 seconds, then centrifuge it for 2 minutes.
d. Verify that the liquid is at the bottom of each well of the plate. If not, centrifuge the plate again
at a higher rpm and for a longer time.
IMPORTANT! Do not allow the bottom of the plate to become dirty. Fluids and other
contaminants that adhere to the bottom of the plate can contaminate the sample block(s) and
cause an abnormally high background signal.
Correct
Liquid is at • Not centrifuged with enough force, or
bottom of well
• Not centrifuged for enough time
To prepare the array card:
a.
Remove the calibration mix from the freezer, then allow it to thaw to room temperature in the
dark (approximately 5 minutes).
IMPORTANT!
Keep the calibration mix out of the light. The fluorescent dye in the mix is
photosensitive. Prolonged exposure to light can diminish the fluorescence of the dye.
b.
Fill the array card with calibration mix. For a normalization calibration, fill two cards with
separate normalization solutions.
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• ViiA
4459846)
• Centrifuge with plate adapter
• Powder-free gloves
• Safety goggles
® ™
/ROX Dyes
Incorrect
Calibrate the ViiA
7 System Fast 96-Well Plate Adapter (PN
™
7 System
3