Applied Biosystems 7900HT Fast Real-Time PCR System and SDS ...

Chapter 7Maintaining the InstrumentPerforming a Pure Dye RunWhen to PerformPurpose of PureDye RunsComponents ofthe Pure DyeSpectraApplied Biosystems recommends performing spectral calibration:• Every 6 months depending on instrument use• After changing sample block formats (see page 7-6)IMPORTANT! Always run a background plate before performing a Pure Dyecalibration.Pure dye data is generated from the results of a pure dye run in which the SDSsoftware collects spectral data from a set of dye standards during a 2-min hold at60 °C. The software stores the spectral information for the pure dye standards in acalibration file located in the SDS directory. After the run, the software extracts eachcomponent dye spectrum from the collected data in the pure spectra run file.IMPORTANT! Because the age and use of instrument components can affect purespectra readings, Applied Biosystems recommends updating the pure spectra datafiles once or twice annually depending on instrument use.The Applied Biosystems 7900HT Fast Real-Time PCR System monitors fluorescentsignals generated by several dyes inside the preloaded pure dye plate. For thestandard 384-well, standard 96-well, or Fast 96-well pure dye plates, the dyes are:FAM , NED , ROX , SYBR ® Green, TAMRA , TET , and VIC ® .For the 7900HT System TaqMan Low Density Array Upgrade, the 7900HTinstrument monitors fluorescent signals generated by three dyes: FAM , VIC ® , andROX . Your installation kit contains these dyes, and you introduce them into threeseparate microfluidic pure dye cards to perform pure dye runs.1 2 3 4 5 6 7DyePeak (nm)1 FAM ~5202 SYBR Green ~5203 TET ~5404 VIC ~550JOE ~5505 NED ~5706 TAMRA ~5807 ROX ~610Figure 7-4Standard Pure Dye Spectra of the 7900HT InstrumentNote: The 7900HT instrument supports the detection of custom pure dyes (dyesother than those provided by Applied Biosystems). To add custom pure dyes to thePure Dye set for your instrument, see “Adding Custom Dyes to the Pure Dye Set” onpage 7-27.7-20 Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User GuideDRAFTSeptember 1, 2004 11:39 am, CH_Maintenance.fm