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Product FAQ

Why are there no RQ values for all the samples in my study from the 7500 or 7500 Fast Real-Time PCR System software?

Answer

The 7500 software requires that the endogenous control be run on every plate within a study. If not, you will only see RQ values for the plate that contains the endogenous control on it. The other plates will have CT values for all the wells, but the software will not calculate RQ values.

If you designed your experiments in this type of layout, you will want to do your analysis in the free DataAssist™ software instead. This program does not have the endogenous control requirement for every plate. Simply export the study results file from the 7500 software as a *.txt or *.csv file, and use that to create a new study in the DataAssist™ software. You should now be able to see RQ values for all samples, even those without an endogenous control on the same plate.

Answer Id: E7241

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Product FAQ

Can I use the 7500 or 7500 Fast Real-Time PCR System to do protein thermal shift experiments?

Answer

Yes. If you have the newer version of the software (v2.0.1 or later), which creates *.eds files, your data will be directly compatible with our Protein Thermal Shift™ Software (https://www.thermofisher.com/order/catalog/product/4466038?ICID=search-product). If you have the older software (v 1.x), you will have to program the software differently (see below) and analyze the results independently. For more details on the analysis, you can refer to this paper: “The use of differential scanning fluorimetry to detect ligand interactions that promote protein stability.” Nat Protoc 2007;2(9):2212-21.

For an experiment using SDS v1.x, follow the directions below:
1.Create a new experiment, choosing Absolute Quantification assay type. Click ‘Next’.
2.Go to ‘Select Detectors’. If there is no predefined detector for this application, create a new one. Make sure the Reporter Dye is ROX, and Quencher Dye is none.
3.Assign the detector to the plate. Select none for the Passive Reference

Answer Id: E7231

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Product FAQ

How can I export or print by Column instead of Row in the instrument software?

Answer

Go to ‘View Well Table’ from the Amp B117Plot screen.

Answer Id: E7232

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Product FAQ

What is the recommended reaction volume that I can run for using 7500 Fast Real-Time PCR System?

Answer

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

Answer Id: E2323

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Product FAQ

Can I collect data at two separate steps for analysis in the 7500 and 7500 Fast Real-Time PCR Systems?

Answer

No. The software will use the last data collection step in the cycling stage for all amplification plots and Ct analysis. So even if you were to set two separate steps with “Data Collection On”, you would only be able to view and analysis data from the latter step.

Answer Id: E7228

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Product FAQ

To analyze an RQ Study, can I import RQ plates from a combination of Fast and Standard mode runs on a 7500 Fast Real-Time PCR System?

Answer

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.

Answer Id: E2319

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Product FAQ

What are the ramp rates for the Applied Biosystems™ 7500 and 7500 Fast Real-Time PCR Instruments?

Answer

The sample ramp rates for the 7500 instrument are as follows:

- Standard Mode: 1.6 deg C/sec up and 1.6 deg C/sec down
- 9600 emulation Mode: 0.8 deg C/sec up and 1.6 deg C/sec down. This matches the sample ramp rate achieved for the ABI PRISM™ 7700 Sequence Detection System.
- Fast Mode: 3.5 deg C/sec up and 3.5 deg C/sec down.

Answer Id: E2481

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Product FAQ

I lost my user manuals for the 7300 and 7500 Fast Real-Time PCR Systems. How can I get an additional copy?

Answer

Please contact Technical Support for Real-Time PCR, and provide the serial number of your instrument. We will email you a copy of the user manual upon request.

Answer Id: E2470

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Product FAQ

What can I use for data analysis?

Answer

Click on the ‘Group By’ button and choose ‘Well Position’ (Column). Then highlight the plate and print the report.

Answer Id: E7233

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Product FAQ

When do I need to calibrate the 7500 or 7500 Fast Real Time PCR System?

Answer

Recommended Maintenance Schedule

Power on/off the computer controlling the instrument: Weekly
Check computer disk space. If necessary, archive or back up your experiment files and instrument settings: Weekly
Background calibration: Every month
Run disk cleanup and disk defragmentation: Every month
Perform an instrument self test: Every month
Pure dye calibrations: Every 6 months
ROI calibration: Every 6 months
Optical calibration: Every 6 months
RNaseP instrument verification: After the instrument has been moved, or as needed to verify instrument performance

Answer Id: E7219

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Product FAQ

Where are my files saved in the 7500 and 7500 Fast Real-Time PCR System software?

Answer

Run files will be saved to a default folder on the connected computer, unless you change it. To find or change the default folder (in SDS v2.0.1 or later), go to Tools then Preferences then Defaults. Here you will see a Data Folder and an Import Folder. The default location is shown. If you want files to be saved to (or open from) a different location, click ‘Browse’ and choose the new folder.

Answer Id: E7229

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Product FAQ

What is the maximum thermal output for the Applied Biosystems™ 7500 / 7500 Fast Sequence Detection System in BTU/h?

Answer

The maximum thermal output for the Applied Biosystems™ 7500/7500 Fast Sequence Detection System is 3241.5 BTU/h (950W).

Answer Id: E2503

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Product FAQ

What are the extensions of files generated by the ABI PRISM™ 7000 Sequence Detection System, Applied Biosystems™ 7300, 7500, 7500Fast, 7900HT Fast Real-Time PCR System, StepOnePlus™ Real-Time PCR System, and ViiA™ 7 Real-Time PCR Systems?

Answer

All run files have an .SDS or .EDS extension. A Relative Quantification study will have an .SDM or .EDM extension and a template will have an .SDT or .EDT extension. The software for StepOnePlus™, ViiA™ 7, and 7500 2.0.x software have .EDS, .EDT, or .EDM extensions; all other software have .SDS, .SDT, or .SDM extensions.

Answer Id: E2473

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Product FAQ

Can I move the Applied Biosystems™ 7300, 7500, or 7500Fast Real-Time PCR System to another room or another building by myself?

Answer

Yes, you can move the instrument carefully on your own. It is recommended that you recalibrate the instrument with the pure dye calibration kit(P/N 4349182, 4349180, 4351151, 4360788, or 4362201) and run an RNase P instrument verification plate (P/N 4351979 or 4350584), after the move.

Answer Id: E2476

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Product FAQ

What should I do if I get “Fatal Error 1420” in SDS 2.0.x software on the 7500 or 7500 Fast Real-Time PCR System?

Answer

You can also Export Data, and the column format will be retained.

Answer Id: E7234

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