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

Chapter 3Preparing a RunRelativeQuantificationWorkflow1. Create a relative quantification plate document (see page 3-9).2. * Apply detectors to the plate document:a. Create detectors for the relative quantification probes (see page 3-11).b. Copy the detectors to the plate document (see page 3-13).3. * Configure the plate document with detector tasks (NTC and Unknown)(see page 3-16).4. * Program the method for the relative quantification run (see page 3-19).5. Choose from the following:– If running a single plate, then continue to step 2.– If running the first plate in a series of plates with identical assayconfigurations, then save the plate document as a template (see page 3-24).6. Create a plate document from the template created in step 2 (see page 3-26).7. Configure the document with sample names and plate information (seepage 3-28).8. Prepare and run the relative quantification plate or plates (see page 4-1).9. Analyze the run data (see page 6-15).*Steps 2 and 2 can be eliminated by importing the plate document setup informationfrom a tab-delimited text file. See “Importing Plate Document Setup Table Files”on page A-2 for more information.Dissociation(Melting) CurveWorkflowThe SDS software can perform a dissociation curve analysis as part of an absolutequantification run. Therefore, to perform a dissociation curve, construct a platedocument for absolute quantification as explained on page 3-5 and configure themethod with a temperature ramp as explained on page 3-23.3-6 Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User GuideDRAFTSeptember 1, 2004 11:39 am, CH_Run.fm