Specifications 12905B24Q0224.pdf

Salient Characteristics of real-time PCR machine

RT-PCR machine must generate a thermal gradient across the 96-well reaction block to support rapid optimization of real-time PCR assays. It must be compatible with many fluorescent technologies for real-time PCR amplification. It needs to capture up to five fluorophores in a single reaction well. It should include software that supports data acquisition, viewing, and analysis of data generated by the system and that supports real-time PCR applications such as PCR quantification, melt curve analysis, end point analysis, allelic discrimination, and gene expression analysis.

Detailed requirements:

1. Real-time data traces must be able to be viewed during a run from the thermal cycler base or if a computer is connected to the instrument.

2. System must be able to detect a 1.3-fold change in gene expression.

3. System must be capable of a thermal gradient that enables optimization of multiple temperatures in a single assay.

4. Thermal block must have operational range of 4–100°C.

5. Sample block temperature must have accuracy of +/-0.2°C of programmed target at 90°C, with a uniformity of +/-0.3°C well-to-well within 10 seconds of arrival at 90°C.

6. An optical system that allows excitation and detection of up to five fluorescent dyes in a single reaction well.

7. Must have a fixed optical path, directly over each well, that eliminates the need to normalize for positional bias.

8. Optical detection range of excitation wavelengths: 450–684 nm;

9. Optical detection range of emission wavelengths: 515–730 nm

10. Must have a linear dynamic range of 10 orders of magnitude for detection of input samples.

11. System capable of detecting one copy of target sequence in genomic DNA.

12. Be able to use reaction volumes from 1–50 µL.

13. Must be able to detect 10 fmol of fluorescein.

14. Instrument must be capable of continuing and completing a run if the software is stopped, or interrupted to prevent loss of data due to unintentional interruption.

15. Instrument must use a Peltier heating/cooling system;

16. Physical Dimensions should be in the range of (W x D x H) 13” x 22” x 14”;

17. Weight should not exceed 48 pounds

Additional software requirements:

18. Multiplex amplification and melt-curve/end-point analysis can be performed with up to five fluorophores in a single well;

19. Multiple plates can be combined into a single experiment;

20. Multiple standard curves can be simultaneously viewed when using a common fluorophore;

21. Gene expression analysis can be completed using multiple motifs (ΔCq, ΔΔCq, etc…)

22. Multiple reference genes can be used in expression analysis,

23. Can compare up to 5000 Cq values

24. Data/graphs/tables MUST be able to be printed, saved as PDF format, or exported directly into

Microsoft Excel;

25. Software must be capable of performing t-test and ANOVA calculations;

26. Software must be capable of displaying data in multiple formats including bar chart, dot plot, scatter plot, etc… and allow IN SITU annotation of those graphs.

27. Software on one computer must be able to directly control up to RT-PCR instruments

28. Multiplex data can be displayed for up to five fluorophores.