Applied Biosystems 7500 - Installing the Software and Powering On the Instrument; Installing the SDS Software for the 7300;7500;7500 Fast System; Powering On the Instrument; Performing the Instrument Function Test

Chapter 5 Generating Data from RQ Plates – 7500 Fast System
Preparing the PCR Master Mix
Applied Biosystems 7300/7500/7500 Fast Real-Time PCR System Relative Quantification Getting Started Guide 39
Notes
FAST
FAST
FAST
5
Probes and primers you design using Primer Express software, must be optimized to
work with the universal assay conditions, using the volumes listed in the following table.
All TaqMan
®
Custom Gene Expression Assays and TaqMan
®
Gene Expression Assays
are formulated so that the final concentration of the primers and probes are within the
recommended values.
Reaction Component µL/ Sample Final Concentration
Taq Ma n
®
Fast Universal PCR Master
Mix (2✕)
10.0 1✕
Forward primer 2.0 50 to 900 nM
Reverse primer 2.0 50 to 900 nM
Taq Ma n
®
probe 2.0 50 to 250 nM
cDNA sample 2.0 10 to 100 ng
Nuclease-free water 20 —
Tot a l 20 .0 —