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

TaqMan Low Density ArrayTable 8-6Troubleshooting the TaqMan ® Low Density ArrayObservation Possible Cause Recommended ActionAfter pipetting, too littlePCR reaction mixture hasgone into the fill reservoir.After pipetting, some of thePCR reaction mixture leaksout of the vent port in thefill reservoir.The PCR reaction mixtureleaks during PCR cycling.After a mean assay resultand standard deviation ofthe replicates arecalculated, a specificreplicate appears to be anoutlier.The PCR reaction mixturewas not correctly pipettedinto the fill reservoir.The aluminum foil backingmay have been damagedduring the sealing step.N/ACare must be taken tocorrectly pipette the PCRreaction mixture (100 µL)into the fill reservoir.1. Carefully follow theprocedures provided in“Loading the TaqManLow Density Arrays” onpage 4-14.2. After centrifuging, besure to note the fluidlevels in the fill reservoiras described in step 6 onpage 4-18.Discard the Low DensityArray.Note: See “About theSealer” on page 4-14 forproper sealing instructions.Review the multicomponentanalysis display for thatreplicate. Delete the outlierand reanalyze.Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User Guide 8-19DRAFTSeptember 1, 2004 11:39 am, CH_Trouble.fm