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

Step 5 – Programming the Plate Document MethodProgramming aTemperatureRamp forDissociationCurve AnalysisTo generate the data required to perform a dissociation curve analysis, the 7900HTinstrument must be programmed to run a ‘temperature ramp’ in which it slowlyelevates the temperature of the samples while collecting fluorescence measurementsonce every 7-10 seconds (see page 6-38 for a detailed explanation).1. In the Instrument tab of the plate document, select the Thermal Profile tab.2. Click the step to the left of the location you want to place the new stage.3. In the Thermal Profile tab, click .The SDS software inserts a temperature ramp at the end of the thermal profileconsisting of a set of default steps.4. You can customize the default temperature ramp, however Applied Biosystemsrecommends that you observe the following guidelines to ensure the maximumresolution for the run (the greatest separation of the derivative peaks during theanalysis).GuidelineExampleThe Start and End steps ofthe temperature ramp must:• be separated by aminimum temperaturedifference of 35 °C.• elapse 15 seconds (0:15)each.End stepStart stepThe ramp rate setting for theEnd step of the temperatureramp must be 2 %.IMPORTANT! You mustdeselect the 9600 Emulationcheck box to modify theRamp Rate Settings.End stepramp ratesettingThe temperature ramp in thethermal profile must containa data collection icon.Datacollectionicon5. Go to “Step 6 – Saving the Plate Document as a Template” on page 3-24.Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User Guide 3-23DRAFTSeptember 1, 2004 11:39 am, CH_Run.fm