I lost my user manuals for the 7300 and 7500 Fast Real-Time PCR Systems. How can I get an additional copy?
What dyes can be used as a reporter in a multiplex reaction on Applied Biosystems® 7500 Real-Time PCR System?
The following dyes can be used as the reporter dye with MGB-NFQ as the quencher on the Applied Biosystems® 7500 Real-time PCR System or Applied Biosystems® 7500 Fast Real-Time PCR System when using TaqMan Gene Expression Master Mix (recommended for multiplexing) or any other Applied Biosystems® TaqMan® master mix:
FAM™, VIC®, JOE™, NED™, TAMRA™,CY3®, ROX™, TEXAS RED®, CY5® dyes.
Of these dyes, we recommend a duplex reaction with FAM™ and VIC® or FAM™ and NED™ dyes; if you are triplexing, we recommend FAM™, VIC®, and Cy5® or FAM™, VIC®, and TAMRA™ dyes. You must choose dyes that are well separated in emission wavelength. If you are using any probes with a TAMRA™ dye quencher, you cannot use NED™, TAMRA™, or CY3® dye as one of the reporter dyes in that multiplex reaction. ROX™ or TEXAS RED® dyes can be used as reporter dyes only when not using an Applied Biosystems® Master Mix.
Answer Id: 2472
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?
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: 2473
How can I run a standard TaqMan® assay reaction on the Applied Biosystems® 7500 Fast Real-Time PCR System?
You can run an Applied Biosystems® 7500 Fast Real-Time PCR System instrument in Standard mode as long the following set-up points are followed:
1) The reaction volume is between 10 ul-30 ul
2) The 96-well plate is a MicroAmp® FAST 96 well reaction plate (P/N 4346907, 4346906 or 4366932)
3) Select the Run Mode to be “Standard 7500 “or “9600 Emulation” (2.0.x software versions only offer Standard or Fast - no 9600 emulation choice).
Answer Id: 2474
Can I move the Applied Biosystems® 7300, 7500, or 7500Fast Real-Time PCR System to another room or another building by myself?
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: 2476
How can I export the amplification plot or gene expression plot from the Applied Biosystems® 7300, 7500, and 7500 Fast Real-Time PCR System’s software?
With SDS 1.4 software, right-click on the graph, and choose "Export as JPEG" or "Export to Powerpoint". With SDS 2.0.x (for 7500/7500fast only), you can either click on the save icon above the graph "Save as JPEG Image" or click on the Export drop-down arrow (located on the upper toolbar) and choose "Send to Powerpoint"
Answer Id: 2477
What are the ramp rates for the Applied Biosystems® 7500 and 7500 Fast Real-Time PCR Instruments?
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: 2481
Can I run "standard" and "9600 emulation" modes on the 7500 Fast Real-Time PCR System?
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.
Answer Id: 2318
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?
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: 2319
What is the recommended reaction volume that I can run for using 7500 Fast Real-Time PCR System?
Which computer operating systems are compatible with the7500 Fast Real-Time PCR System?
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.
Answer Id: 2327
If I were to upgrade my existing 7500 System standard unit to the a 7500 Fast configuration, can I still open my old data files?
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.
Answer Id: 2341
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.
Answer Id: 2342
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.
Answer Id: 2343
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.
Answer Id: 2344