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

Chapter 4Operating the InstrumentPreventing ContaminationContaminationand the5´ NucleaseAssayGeneral PCRPracticesPCR using the 5´ nuclease assay requires special laboratory practices to avoid falsepositive amplifications (Kwok and Higuchi, 1989). The assay’s logarithmicamplifications can potentially amplify a single DNA molecule (Saiki et al., 1985;Mullis and Faloona, 1987).Observe the following guidelines when assembling optical plates or loadingTaqMan ® Low Density Arrays for use on the 7900HT instrument:• Wear a clean lab coat (not previously worn while handling amplified PCR productsor used during sample preparation) and clean gloves when preparing samples forPCR amplification.• Change gloves whenever you suspect they are contaminated.• Maintain separate areas, dedicated equipment, and supplies for:– Sample preparation– PCR setup– PCR amplification– Analysis of PCR products• Never bring amplified PCR products into the PCR setup area.• Open and close all sample tubes carefully. Try not to splash or spray PCR samples.• Keep reactions and components capped as much as possible.• Use positive-displacement pipets or aerosol-resistant pipet tips.• Clean lab benches and equipment with 10% bleach solution.Compatible ConsumablesConsumables forUse withthe 7900HTInstrumentTable 4-1 Compatible ConsumablesConsumable TypeUse these…Compatible SealsNot these…ABI PRISM 384-Well OpticalReaction Plates• ABI PRISM Optical Adhesive Covers(quantity 100)• Optical Adhesive Covers (quantity 25)384-Well Septa4-6 Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User GuideDRAFTSeptember 1, 2004 11:39 am, CH_Basic.fm