DRAFT
September 1, 2004 11:39 am, CH_Maintenance.fm
Performing a Pure Dye Run
Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User Guide 7-21
After a run, the SDS software receives run data in the form of a raw spectra signal for
each reading. To make sense of the raw data, the software must determine the
contribution of each fluorescent dye used in the sample through a process called
multicomponenting. The software accomplishes the separation by comparing the raw
spectra with a set of pure dye standards contained in a calibration file. When a plate
document is saved after analysis, the software stores the pure spectra information
with the collected fluorescent data for that experiment.
Materials
Required
384- or 96-Well Plate
• Appropriate Spectral Calibration Kit:
– Sequence Detection Systems 384-Well Spectral Calibration Kit
–ABIP
RISM
â„¢
7900HT Sequence Detection Systems 96-Well Spectral
Calibration Kit
– 7900HT System Fast 96-Well Spectral Calibration Kit
Note: Both the standard 96-well and Fast 96-Well Spectral Calibration Kits
contain two pure dye plates.
• Product Insert
• Centrifuge, with plate adapter
Microfluidic Pure Dye Cards
• Microfluidic card spectral calibration reagents
• Centrifuge
• Microfluidic card sealer