Notebook Computer Kit for 7300/7500/7500 Fast Real-Time PCR Systems - FAQs

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.

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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.

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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.

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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.

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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.

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Yes, older 7500 or 7500 fast data files are compatible with the current version of the SDS software. However, there is one exception: please note that the SDS v1.3 software will not allow RQ Studies to be created from RQ Plates from different instrument types, different thermal modes (i.e., fast and standard modes) or different numbers of thermal cycles.

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You can use up to Windows XP, Service Pack 3 (SP3) on the computer that is connected to the 7500 Fast Real-Time PCR System instrument. An exception is the SDS v2.0.5 software, which can be used with the Windows 7 operating system for stand-alone data analysis steps.

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The recommended input template range is 10-100 ng of gDNA/ cDNA template per reaction.

We recommend running a total reaction volume between 10 and 30 ul per well.

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We do not recommend this as the current and previous versions of SDS software will not allow RQ Studies to be created from RQ Plates from different instrument types, different thermal modes/profiles or different numbers of thermal cycles. It is important to reduce variability among RQ plates in an RQ study by importing RQ plates from the same instrument type, with the same thermal profile and mode, as well as the same block type and sample volume.

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Yes, standard and 9600 emulation modes are optionally available. We have matched the ramping rate and thus anticipate equivalent performance. However, the actual total run times will differ from standard and 9600 emulation modes run on a standard block, and we cannot guarantee identical results as performance may vary depending on the assay.

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Yes, the standard 7500 instrument has a 96 x 0.2 mL block, whereas the 7500 Fast instrument has a 96 x 0.1 mL block.

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