Appendix B
Applied Biosystems 7300/7500/7500 Fast Real-Time PCR System Installation and Maintenance Guide 125
Notes
B
Creating a Background Plate
Overview
Whenever possible, use a background plate included with the spectral calibration kit.
The plates supplied in the kit contain a buffer that accurately models the reagents used
for PCR, and, therefore, produces high-quality calibration data. However, if a
background plate from a spectral calibration kit is not available, you can create one by
following the procedure below.
Materials
Required
Creating a Background Plate
IMPORTANT! Wear powder-free gloves while creating the background
plate.
1. Remove an Applied Biosystems 96-Well Optical Reaction Plate or 96-Well Fast
Reaction Plate from its box and place it on a clean, dry surface.
2. Aliquot 50 µL (7300/7500 system) or 20 µL (7500 Fast system) of deionized water
to each well of the reaction plate.
3. Seal the plate using an optical adhesive cover or optical flat caps.
Use the plate for background calibration in the same way you use a background plate
from the spectral calibration kit. See Chapter 5, “Performing the Background Calibration
and Optical Calibration.”
Applied Biosystems
Optical 96-Well
Reaction Plate or 96-
Well Fast Reaction
Optical Adhesive Cover
or Optical Flat Caps
Deionized
water
Pipettor, 200-µL
(with pipette tips)
Safety glasses
Powder-free
gloves
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