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

Chapter 4Operating the InstrumentPreparing Optical Plates for UsePreparing Platesfor Use on the7900HTInstrument1. Prepare the reactions in an optical plate by aliquoting reagents, enzyme, andsamples to the appropriate wells of an optical plate.IMPORTANT! The arrangement of the reactions (samples and assays) on theplate must match the configuration of information (sample names anddetectors/markers) on the corresponding plate document.2. Seal the optical plate with a seal that is compatible with the 7900HT instrument(Figures 4-3 and 4-4 on page 4-9).Note: See “Compatible Consumables” on page 4-6 for a list of seals for usewith the Applied Biosystems 7900HT Fast Real-Time PCR System.3. Briefly centrifuge the plate to collect the reactions at the bottom of the wells andto eliminate any air bubbles that may be present.4. Visually verify that each reaction is positioned at the bottom of its well.Correct PositionIncorrect PositionThe sample is positionedcorrectly in the bottom ofthe well.The sample lies on theside wall because theplate was not centrifuged.An air bubble lies at thebottom of the wellbecause the plate wasnot:• Centrifuged with enough force, or• Centrifuged for enough time5. If running an ABI PRISM 96-Well Optical Reaction Plate, apply the appropriatecompression pad to the sealed optical plate.If you are going to run the plate:• Individually using the SDS software, then apply a standard compressionpad (Figure 4-3)• As part of a batch using the Automation Controller Software, then apply anABI PRISM Snap-On Compression Pad (Figure 4-3)IMPORTANT! The ABI PRISM Snap-On Compression Pad/plate assemblymay still be hot after the Zymark ® Twister Microplate Handler loads it intothe Plate Handler’s output stack. Please wait at least 30 seconds beforemanually handling the Snap-On Compression Pad/plate assembly.6. Choose from the following. If performing:– Absolute or Relative Quantification, or Dissociation Curve AnalysisRun the plate as explained in “Running the Plate” on page 4-25.– Allelic Discrimination4-8 Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User GuideDRAFTSeptember 1, 2004 11:39 am, CH_Basic.fm