Spartan DX-12 FAQ
| Q30. | Are the Spartan DXTM & Spartan DX-12TMlicensed for real-time PCR? |
| Q31. | Is the Spartan DX-12TMlicensed for melt-curve applications? |
Answers Section
Spartan DX-12TM Instrument
Q1. What are the excitation and emission wavelengths for the 2 channels of the Spartan DX-12TM?
A1: The FAM channel has an excitation wavelength of 471-492 nm and an emission wavelength of 520-532 nm. The Texas-Red channel has an excitation wavelength of 555-584 nm and an emission wavelength of 620-800 nm.
Q2. Does the Spartan DX-12TM have a heated lid?
A2: No, the Spartan DX-12TM does not have a heated lid. The Spartan tube and cap are designed for even heat distribution and eliminate the requirement for oil or other sealants.
Q3. What is the technology behind the Spartan DX-12TM’s heating and cooling system?
A3: The Spartan DX-12TM uses a patented technology with fixed-temperature thermal blocks.
Q4. How uniform is the temperature across the 12 wells of the Spartan DX-12TM?
A4: The inter-well temperature is +- 0.5C at steady state between 40C and 95C.
Q5. What reaction volumes can be used in the Spartan DX-12TM?
A5: The Spartan DX-12TM tube is designed for 10-20 ul reaction volumes. Reaction volumes less than 20 ul are suitable for qualitative applications, but we recommend 20 ul for most applications.
Q6. Does the Spartan DX-12TM have gradient capability?
A6: No, the instrument does not have gradient capability.
Q7. Does the Spartan DX-12TM require a reference dye like ROX?
A7: No, the instrument does not require a reference dye.
Q8. Is it possible for the user to run reactions using ROX dye as passive reference dye? Can the Spartan Graphing and Analysis software normalize to a ROX signal?
A8: Although the instrument does not require a reference dye such as ROX, users can run kits or homebrew reactions containing ROX as a passive reference dye. However, the ROX data is not required nor used for normalization.
Q9. What is the detection sensitivity of the Spartan DX-12TM?
A9: The sensitivity of an assay is affected by a number of factors including: (1) the level of background fluorescence, (2) the formation of non-specific prvoducts, and (3) the concentration of the probe and the presence of inhibitors. The Spartan DX-12TM is able to detect <10 copies of DNA for properly-designed PCR assays, with appropriate fluorescence technology.
Q10. How are the fluorescence measurements taken for the 12 wells of the Spartan DX-12TM?
A10: The data from all 12 wells of the Spartan DX-12TM is collected simultaneously.
Q11. Can the excitation and emission filters be changed by the user?
A11: No, the filters in the instrument cannot be changed by the user. The instrument’s analysis algorithm is designed for dyes in the specified optical ranges.
Q12. Can the Spartan DX-12TM perform a reverse transcription step?
A12: Yes.
Q13. How is melt curve data acquired by the Spartan DX-12TM?
A13: During melt-curve analysis, the Spartan DX-12TM employs a linear thermal ramp that acquires data at increments between 0.1C and 1C/reading.
Q14. What type optical detection technology does the Spartan DX-12TM use?
A14: The Spartan DX-12TM uses a 10-bit CMOS camera to accurately capture and record fluorescence data. An on-board analysis algorithm adjusts and normalizes the data to correct for fluorescence cross-talk and background.
Q15. What should I do if I have concerns about my Spartan DX-12TM’s performance?
A15: Please refer to our troubleshooting guide (www.spartanbio.com/support/troubleshooting)
Spartan Graphing & Analysis Software
Q16. Is the Spartan Graphing & Analysis software provided free of charge?
A16: Yes, the Spartan Graphing & Analysis software comes free of charge with the purchase of a Spartan DX-12TM instrument. The end-user licence agreement must be accepted before the software can be installed.
Q17. What are the minimum computer system specifications required for running the Spartan Graphing & Analysis software?
A17: Spartan Bioscience recommends the following minimum requirements for your computer system:
Pentium IV processor
USB port (1.1 or 2.0)
500 MHz
20 GB available on hard drive
512 MB of RAM
Windows XP operating system
UL 60950-1 listed laptop or desktop computer
Q18. How do I transfer saved data from the Spartan DX-12TM to my PC?
A18: When the instrument is finished saving the run data, transfer your data by disconnecting the USB drive from the instrument and connecting it to your PC. Open the Spartan Graphing & Analysis software and press ‘Ctrl+I’. Select the new file from the USB directory and click ‘Open’. The file can be found in the ‘Run data/unit’ directory and identified by the naming convention AAAAAAA_UUUU_DD_MM_RR.txt, where AAAAAAA is the program name, UUUU is the unit ID number, DDMM are day and month, and RR is the run of the day. For more information, consult ‘Section 3. Basic Operations’ in the Spartan Graphing & Analysis Software Manual.
Q19. When do I input the run information on the Spartan-DX-12TM?
A19: The Spartan DX-12TM does not require the input of run information prior to starting PCR. The run information is usually entered into the ‘Run Data’ tab of the Spartan Graphing and Analysis Software once the run is completed. Alternatively, the run information can be entered into a template that is then applied to the data after it has been loaded in the Spartan Graphing and Analysis software.
Q20. Can users be prevented from modifying or deleting important templates?
A20: The Spartan Graphing and Analysis software has a password-protected ‘Administrator Mode’. Turning on this mode prevents non-administrators from accidentally modifying existing templates, analysis methods, and plus/minus result-calling parameters. Non-administrators are still able to create their own unlocked templates and files.
Q21. Does the Spartan Graphing & Analysis software allow for the rearrangement of the samples in the graphical representation?
A21: The software enables users to select/deselect which wells are presented in the graphs and/or selected for the generation of a standard curve.
Q22. Can I use the Spartan Graphing & Analysis software to analyze data from both the Spartan DX-12TM and as well as the original Spartan DXTM?
A22: The Spartan Graphing & Analysis software is only compatible with the Spartan DX-12TM. For a compatible version of the software for the Spartan DXTM, please contact support@spartanbio.com.
Q23. Can graphs from the Spartan Graphing & Analysis software be copied into Microsoft Excel, Word or PowerPoint files?
A23: Yes. All graphs may be saved as image files by right-clicking on the graph and copying it to the clipboard or by exporting them as JPG files. To insert the image into Microsoft Excel, Word, or PowerPoint, first copy the image by selecting Edit/Copy (Ctrl+C), and then insert the image by selecting Edit/Paste (Ctrl+V).
Data Analysis
Q24. Can the Spartan Graphing & Analysis Software perform absolute and relative quantification?
A24: Yes, the Spartan Graphing & Analysis software is designed to perform absolute and relative quantification using standard curves. Standard curves may be exported from one experiment, and then imported into another experiment and normalized to a reference standard. Alternatively, Crossing Threshold (Ct) data may be exported and then analyzed using methods such as delta-delta-Ct (??Ct).
Q25. Can the Spartan Graphing & Analysis Software perform plus/minus calling (results calling)?
A25: Yes, the software may be set up to do plus/minus or results calling based on parameters determined by an administrator. This can be done through the Template Setup Menu. For details see Section 8 of the Spartan Graphing & Analysis Software Manual.
Q26. Can the user export raw data into Microsoft Excel to do further analysis, or to manually analyse the data?
A26: Yes. In the Spartan Graphing & Analysis software, export the data as a ‘*.txt’ file by selecting ‘File/Export Raw Data’ and then saving the file to the desired location. This .txt file may then be imported into Microsoft ExcelTM.
Q27. How does the Spartan DX-12TM separate the different fluorophore signals?
A27: The Spartan DX-12TM utilizes a multi-component algorithm to deconvolve the fluorescence signals acquired by the camera.
Q28. How is the Threshold Cycle (Ct) value selected?
A28: The Spartan Graphing & Analysis software identifies the Ct as the point on the real-time data curve where acceleration of the fluorescence signal reaches a maximum.
Q29. Can a reference point across multiple curves be selected manually by the user?
A29: The Spartan Graphing & Analysis software allows the user to choose between ‘Linear Fit’ or ‘log Fit’ calculation methods. In the ‘Linear Fit’ mode, the Ct is determined automatically. In the ‘Log Fit’ mode, the user can manually set a crossing point (CP) by adjusting the horizontal CP line. CP thresholds should be chosen within the log-linear region of all real-time curves. For more information, consult Section 4.10 of the Spartan Graphing & Analysis Software Manual.
Licensing
Q30. Are the Spartan DXTM & Spartan DX-12TM licensed for real-time PCR?
A30: No, neither the Spartan DXTM nor the Spartan DX-12TM are licensed for real-time PCR. The default setting for both instruments is end-point detection. In the Options Menu, the real-time detection setting is limited to users who either have a license or do not require a license to use real-time PCR technology.
Q31. Is the Spartan DX-12TM licensed for melt-curve applications?
A31: No, the Spartan DX-12TM is not licensed for Melt-curve applications. The default setting for the Spartan DX-12TM is melt-curve functionality ‘Off’. In the Options Menu, the Melt-curve ‘On’ setting is limited to users who either have a license or do not require a license for melt curve applications.
Chemistry
Q32. Can fluorescing quenchers be used with the Spartan DX-12TM?
A32: Yes, fluorescing quenchers such as TAMRA and Dabcyl may be used. Note that better performance may be achieved by using non-fluorescing quenchers such as Black Hole QuenchersTM (BHQ) or Non-fluorescing Quenchers (NFQ).
Q33. Can MGBTM, LNATM or BHQPlusTM probes be used with the Spartan DX-12TM?
A33: Yes, MGBTM, LNATM and BHQPlusTM probes may be used on the Spartan DX-12TM.
Q34. Can commercial kits be run with the Spartan DX-12TM?
A34: Yes, commercial kits may be run on the Spartan DX-12TM, provided that they have fluorophore/quencher combinations which are compatible with the instrument’s 2 optical channels.
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