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Applied Biosystems™ StepOne™ Real-Time PCR System, laptop

Catalog No. 4376373
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Catalog No. 4376373 Supplier Applied Biosystems™ Supplier No. 4376373

Please call Customer Service at 1-800-234-7437 or send an email to help@thermofisher.com for assistance.

Description

Includes

Laptop Computer

StepOne™ Real-Time PCR System is a 48-well, low-throughput Real-Time PCR instrument perfect for both first-time and experienced users.

The StepOne™ Real-Time PCR System can be setup in a variety of configurations and comes ready to use, out of the box, with intuitive data analysis and instrument control software. Utilizing robust LED based 3-color optical recording, the StepOne™ Real-Time PCR System is designed to deliver precise, quantitative Real-Time PCR results for a variety of genomic research applications. Includes a laptop computer.

Features of the The StepOne™ Real-Time PCR System include:

  • Advanced software and instrumentation for performing a wide array of genomic assays.
  • Sensitive 3-color optical LED recording syste
  • Intuitive and robust software perfect for both first-time and advanced users
  • Simple and flexible instrument set-up and usage

StepOne™ Software:

The StepOne™ Software included with the StepOne™ Real-Time PCR System runs on Windows™ XP, Window Vista™, and Windows™ 7 operating systems and provides instrument control, data collection, and data analysis capabilities. This latest version includes capabilities to collect melt curve data for High Resolution Melt (HRM) applications and the option to export in Real-Time PCR Data Markup Language (RDML) for compatibility with MIQE guidelines.

Rapid and Simple Assay Set-Up:

This remarkably simple Real-Time PCR system is designed with an LCD touchscreen and a user-friendly interface that brings the power of genetic studies to researchers new to Real-Time PCR. The intuitive software and protocol wizards help guide new users through their Real-Time PCR experiments. The StepOne™ Real-Time PCR System also includes a quick-start setup so that you start a run immediately and enter plate information at a later time.

Sensitive LED-Based 3-Color Fluorescence Reading:

The StepOne™ Real-Time PCR System utilizes a long-life LED-based optical system that can record fluorescence from FAM™/SYBR™ Green, VIC™/JOE™, and ROX™ dyes. This cost-effective, 3-color, 48-well system delivers precise, quantitative Real-Time PCR results and saves data from all filters in every run without depending on a computer or plate setup. It can discriminate between 2 populations of 5,000 and 10,000 template copies of a TaqMan™ assay with 99.7% confidence.

Compatible with Many Genomic Analysis Techniques:

Perform a variety of standard and demanding genetic analysis research techniques on one instrument using the user friendly StepOne™ Real-Time PCR System and software. The StepOne™ System supports many Real-Time PCR applications, including the following:

  • SNP Genotyping
  • Gene Expression Analysis
  • MicroRNA Expression
  • Translocation Analysis
  • Gene Detection
  • Viral Load Analysis
The included StepOne™ Software supports a variety of analysis methods, including the following:
  • Standard Curve (absolute quantitation)
  • Relative Standard Curve
  • Comparative Ct (ΔΔCt) (relative quantitation)
  • Genotyping and Presence/Absence
  • Melt Curve Analysis
  • High Resolution Melting (additional software required)

Versatile Instrument Configuration Options:

The ultra-compact footprint of a StepOne™ System can be installed in multiple distinct configurations, providing unmatched flexibility and convenience that can allow a fit in any laboratory. The StepOne™ Real-Time PCR System can be used via a PC, with a PC connected to a Local Area Network (LAN), or as a stand-alone instrument (PC-free). Each StepOne™ Real-Time PCR System is factory-calibrated for optical and thermal accuracy, so simply remarkable Real-Time PCR results can be obtained right out of the box.

Real-Time Data Monitoring, Dissemination, and Storage:

The system measures amplification as it occurs, allowing you to monitor the progress of your experiment cycle by cycle, either on the machine or remotely. Your data is stored on the instrument itself, and can be viewed and stored automatically via remote access or transferred via email or a USB flash drive. Data can also be conveniently exported as PowerPoint™, Excel™, and graphical image files.

Expanded Gene Expression Capabilities:

The advanced software provided with the StepOne™ Real-Time PCR System now includes the powerful Gene Expression Study Package. This software package allows for greater flexibility and accuracy in your gene expression assays through:

  • Analysis of an unlimited number of plates in one study
  • Sorting of data by biological or technical replicate group
  • Use of multiple endogenous controls
  • Assay efficiency correction

High Resolution Melting Capability:

Now with High Resolution Melt Software v3.0 you can perform post-PCR sequence variation analysis with the StepOne™ Real-Time PCR System. This separately purchased software simplifies set-up by maintaining assay specific settings and accepts pasted plate layout information directly from Excel. HRM Software v3.0 also has an improved clustering algorithm for increased sensitivity and accuracy, and the ability to conduct separate analyses of multiple assays run on a single plate.

For Research Use Only. Not for use in diagnostics procedures.

Specifications

Format 48-well Plate, 0.1 mL Tubes
Thermal Accuracy ± 0.25°C (35°C to 95°C) of Display Temperature
Sample Ramp Rate Fast Mode: ± 2.2°C/s, Standard mode: ± 1.6°C/s
Sensitivity 1 Copy
Temperature Range (Metric) 4 to 100°C
Reaction Volume Range 10–30 μL (Standard Curve Experiments: 40 μL in Standard Mode is Validated)
Passive Reference Dye ROX (Separate Tube), No ROX, ROX (Pre-mixed)
Dynamic Range Linear Dynamic Range greater than 9 Log Units (Detection)
Content And Storage Laptop Computer
Calibrated Dye VIC™, SYBR™ Green I, JOE™, FAM™, ROX™
High-throughput Compatibility Multiplexing
For Use With (Equipment) StepOne™
Thermal Uniformity ± 0.50°C (35°C to 95°C) of Setpoint/Display Temperature
Reaction Speed Fast, Standard
Detection Method SYBR, Primer-Probe Detection
Run Time <2 hrs/run (Standard Mode), <40 min/run (Fast Mode)
Optics LED, 3 Emission Filters, Photodiode
Thermal Cycling System Peltier-based System
Dimensions 24.6 cm/9.7 in.(W) x 42.7 cm/16.8 in.(D) x 51.2 cm/20.2 in.(H)
Capacity 48 x 0.1 mL Tubes, 1 x 48-well Plate
External Computer Notebook
Product Line StepOne
Peak Block Ramp Rate 4.6°C/s
Display Type LCD Touchscreen
Quantity 1 instrument
Weight 23.6 kg (52 lb.)
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Frequently Asked Questions (FAQs)

What does "Error - Cannot calculate Pure Dye Matrix" mean?

This issue can occur if the run is interrupted. To prevent this from happening in the future, avoid ending the PCR with an infinite hold stage or interrupting the run. Please send the file to techsupport@thermofisher.com so that our software group can attempt to recover the data.

What does "Error - Fatal Error Number 0x8158" mean?

This is generally caused by incorrect placement of tube strips, resulting in a sample block that can't properly be lowered. We recommend contacting Instrument Services to have an engineer walk you through a manual drawer-lowering process during instrument power-up. To prevent this from happening in the future, place tubes toward the middle of the block or use a placeholder strip on the opposing side to counterbalance block pressure.

What does "Error - Java.lang.IllegalArgumentException: One of the raw spectra is null" mean?

This issue can occur if the “Start Run” button is clicked more than once or if illegal characters are used in the experiment/file name. You can recover the data yourself by overriding the calibration file with one that represents the calibration status of the instrument when you ran that particular file. Use the toolbar at the top to navigate: Analysis then Override Calibration then Use Calibration From Another File.

What does "Error - Cannot open the file because file contents are not recognized" mean?

This issue can occur if duplicate samples are added to the experiment, if illegal characters are used in the experiment/file name, or if the file extension was changed to .eds from a different extension.
Please send the file to techsupport@thermofisher.com to attempt recovery.

What does "Error - Analysis failed due to GExSession doesn't exist in context" mean?

This error means that the data collection was not turned on as appropriate. If this is inadvertently turned off, no data will be collected. Thus, the data are not retrievable in such a file.

What does "Error - Analysis failed due to: 1" mean on the StepOne or StepOnePlus Real-Time PCR System?

Please send the file to techsupport@thermofisher.com so that we can have our software group attempt to recover the data.

What should I do when I get the error "Error - Cannot connect to the instrument" on the StepOne or StepOnePlus Real-Time PCR System?

Check that all the cables are correctly plugged into the respective ports and not bent. Follow the directions on page 135 of the StepOne and StepOnePlus Maintenance Guide (https://tools.thermofisher.com/content/sfs/manuals/4376782.pdf) to check the connectivity. If the issue is still not resolved, please contact Instrument Services at 1-800-955-6288, option 3, option 1, or by email at instrumentservices@thermofisher.com for further assistance.

What does "Error - Failure to acquire needed controlling rights" mean?

This error usually occurs when you first start your run. Reboot the instrument and restart the computer. Make sure to log into the computer as the Administrator (on the Windows OS).

Where are my files saved to on the StepOne and StepOnePlus Real-Time PCR Systems?

Run files will be saved to both the instrument and the connected computer. On the connected computer, files will be saved to the default data folder, unless you change it.

To find or change the default folder, 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.

How do I set up a touchdown PCR on the StepOne and StepOnePlus Real-Time PCR Systems?

In a touchdown PCR experiment, you will change either the temperature or the time of a particular PCR step with every cycle. Most commonly, the annealing temperature is adjusted throughout the experiment, such that the specificity is higher in the early cycles and the efficiency in the later cycles.

1. Go to File then New Experiment then Advanced Setup. Fill out the relevant options as you normally would.
2. Go to the Run Method under the ‘Setup' section and you should see the Graphical View of your thermal profile. Check the box next to ‘Enable AutoDelta'. You should see some grey triangles appear next to the Temperature and Time at every step in the Cycling Stage. (Note: If you want to start the changes at a later cycle, set this here under ‘Starting Cycles'.)
3. A new window called ‘AutoDelta Settings' will open up. Select the appropriate options and click ‘Save Setting'. You will then see a green triangle show up next to the parameter you changed. Your new method has now been applied.

What plates or tubes can I use with the StepOne and StepOnePlus Real-Time PCR Systems?

Please refer to this selection table (https://www.thermofisher.com/us/en/home/life-science/pcr/pcr-plastics/real-time-pcr-plastics.html) for compatible plates, tubes, and films.

How do I load tube strips on the StepOne and StepOnePlus Real-Time PCR Systems?

Follow these instructions for proper loading of tubes, tube strips, and plates:

- Put the tubes or tube strips in the tray with the adapter on the 96-well support base.
- If using a plate, put the plate directly on the 96-well support base.
- Pipet your reactions into the tubes/tube strips or the wells on the plate.
- Seal the tube/tube strips with optical flat caps.
- If using a plate, seal the plate with either optical adhesive film or flat caps.

For optimal performance on the StepOnePlus instrument with partial loads:
- Load at least 16 tubes and arrange them in adjacent columns of 8 tubes, using rows A through H, or adjacent rows of 8 tubes, using columns 3 through 10.
For optimal performance on the StepOne instrument, load at least 4 tubes in the sample block.

What volumes can be used in my plate for the StepOne or StepOnePlus Real-Time PCR Systems?

The StepOne and StepOnePlus Real-Time PCR Systems have a Fast block, which supports reaction volumes of 10-30 µL.

Can I reuse my calibration plates for the StepOne or StepOnePlus Real-Time PCR Systems?

The calibration plates can be stored and reused three times for up to one year after you receive them, so make sure to return them to their original packaging and return them to -20 degrees C storage until the next use. If needed, you can make your own background plate using deionized water. Please follow the directions in the StepOne and StepOnePlus Maintenance and Administration Guide (Appendix C) (http://tools.thermofisher.com/content/sfs/manuals/4376782G.pdf) for more details.

When do I need to run the RNaseP calibration for the StepOne or StepOnePlus Real-Time PCR Systems?

The RNaseP verification plate is a single-use plate that contains template, master mix, and a TaqMan assay for RNaseP. It is used to verify that the instrument is performing to specifications. If you have reason to suspect that there is something wrong with the instrument, or if you want to rule out a chemistry issue, the RNaseP plate is a good way to test the system. It is also recommended to use after the instrument has been moved to a new location. For the StepOne instrument, the RNaseP plate is Cat. No. 4371439. For the StepOnePlus instrument, the RNaseP plate is Cat. No. 4351979.

When do I need to calibrate the StepOne or StepOnePlus Real-Time PCR System?

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 18 months
Spatial calibration: Every 18 months
RNaseP instrument verification: After the instrument has been moved, or as needed to verify instrument performance

What real-time instruments can be run independently from a computer?

The StepOne, StepOnePlus, ViiA 7, and QuantStudio 12K Flex systems can all be run directly from the instrument touchscreen and don't need a separate computer to operate them.
Can I import a standard curve for my real-time PCR data into the Design & Analysis Software (v2.x)?

It is possible to import a standard curve into the Design & Analysis Software (v2.x) if you are analyzing results from one of the following Applied Biosystems real-time PCR instruments:
• 7900HT Fast Real-Time PCR System
• 7500/7500 Fast Real-Time PCR System
• StepOne/StepOne Plus Real-Time PCR System
• ViiA 7 Real Time PCR System
• QuantStudio 1 Real-Time PCR System
• QuantStudio 3 & 5 Real-Time PCR System
• QuantStudio 6 & 7 Flex Real-Time PCR System
• QuantStudio 6 & 7 Pro Real-Time PCR Systems
• QuantStudio 12K Flex Real-Time PCR system

Follow the instructions below to import a standard curve for analysis in the Design & Analysis Software (v2.x):
1. Open the data file in the Design & Analysis Software (v2.x).
2. Click on Actions.
3. Click on "Standard Curve Analysis Setting".
4. Select "External Standard Curves" and click on "Import".
5. Browse to the proper .csv file and click "Open".
6. Click "Apply".

What is the difference between a Fast 96-well and standard 96-well block?

There are two main differences between a Fast 96-well and standard 96-well block for the Applied Biosystems real-time PCR systems:
• Reaction well volume: The Fast 96-well blocks have 0.1 mL reaction wells and are compatible with 0.1 mL plates and tubes. The standard 96-well blocks have 0.2 mL reaction wells and are compatible with 0.2 mL plates and tubes.
• Ramp rate: The fast 96-well blocks have a higher maximum ramp rate than standard blocks for the same real-time PCR system. However, both the fast 96-well and standard 96-well blocks are capable of running standard and fast chemistry.

Note: For newer instruments (QuantStudio 3 and 5 Real-Time PCR Systems and later) "0.1 mL" and "0.2 mL" are being used in place of "fast" and "standard" as designations for the two 96-well block formats.
What qPCR instruments are compatible with Thermo Fisher Cloud?

All our real-time PCR instruments are compatible with Thermo Fisher Cloud Apps except the 7500 Fast instrument where the eds files alone are compatible with Thermo Fisher Cloud Apps.