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

Calling and Scrutinizing Allelic Discrimination Data3. Screen for Unknown samples that failed to amplify:a. In the Allelic Discrimination Plot, select the NTC cluster.The SDS software highlights the datapoints within the allele plot and theplate grid.b. In the plate grid, check the wells containing Unknown samples for selectedwells that clustered with the NTCs.Samples that clustered with the No Template Control wells may:• Contain no DNA• Contain PCR inhibitors• Be homozygous for a sequence deletionNTC cluster(selected)No TemplateControl wellsUnknown samplesclustered with the NTCs4. Retest any samples that did not cluster tightly or clustered with NTCs toconfirm the results.5. If evaluating for multiple markers, do the following:a. In the Marker drop-down list, select a different marker.b. Repeat steps 1 through 4 for the new marker.c. Repeat steps a and b until the calls for each marker have been verified.Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User Guide 5-17DRAFTSeptember 1, 2004 11:39 am, CH_End-Point.fm