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

Chapter 3Preparing a Run4. Configure the data collection options for the method:a. Select the Data Collection tab.b. Click below each plateau or ramp inside the cycle stage of the thermalprofile to place a data collection icon at each step.Data collection iconsIMPORTANT! Configure the method of your plate document to collect data onlyin the necessary stages of the PCR. The file size for a real-time run depends onthe length of the run and the configuration and placement of the data collectionicons. Longer runs and runs configured to collect data at multiple stages of themethod can be considerably larger than the 15–25 MB average.5. Click the Sample Volume (µL) field and enter the volume of the reactions onthe plate.Note: Sample volume refers to the entire contents of any well, including bufferblank, or any combination of master mix and nucleic acids.IMPORTANT! All wells on one plate must contain the same reaction volume.6. Choose from the following options:If performing an…absolute or relativequantification run onlyabsolute quantification runwith a dissociation curveThen…go to “Step 6 – Saving the Plate Document as aTemplate” on page 3-24.add a temperature ramp to generate dissociationcurve data as explained on page 3-23.3-22 Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User GuideDRAFTSeptember 1, 2004 11:39 am, CH_Run.fm