Can TET dye be used on the Applied Biosystems real-time PCR instruments?
TET dye is not recommended to be used on the following real-time PCR instrument models: 7000, 7300, 7500, 7500 FAST, StepOne, and StepOnePlus Systems.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
What dyes can I use as reporter dyes in a multiplex reaction on the Applied Biosystems 7300 Real-Time PCR Systems?
The Applied Biosystems 7300 Real-Time PCR System has 4 signal detection filters: Filter A (FAM and SYBR Green), Filter B (VIC and JOE), Filter C (TAMRA, NED and Cy3) and Filter D (ROX and Texas Red). For multiplexing reactions, the reporter dyes from the different assays should be chosen from different filter sets, so they can be compatible. In addition, the reporter dye need to be compatible with the quencher and the passive reference dye that might be present in the reaction. The commonly used combination is FAM and VIC.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
What are the thermal cycling parameters of the Fast mode for the 7500 Fast Real-Time PCR System?
For the 7500 Fast real time PCR systems, the thermal cycling parameters are defaulted to 50°C/2 min for UNG incubation, 95°C/20 sec for fast polymerase activation, 40 cycles of 95°C/3 secs for the dissociation and 60°C/30 seconds for annealing and extensions.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
Can I run slower, standard thermal cycling experiments on a 7500 Fast Real-Time PCR System?
Yes. you can choose either standard or fast ramp on the 7500 Fast System for software versions 2x or greater, and a 9600 emulation thermal cycling mode is available for customers who wish to run assays based on adjusted ramp rate. However, you should be aware that we cannot guarantee identical performance to the same modes run on a standard 7500 system block.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
Can I compare the data collected when using Fast thermal cycling mode compared to using standard thermal cycling mode on the 7500 Fast Real-Time PCR System?
Runs can be compared between standard and fast mode, as the data collected are collected in all five filters in the instrument. However, keep in mind that these data may not always show identical results and may have added variability due to the data collection processes and differences in ramp rate between standard and fast mode.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
