Applied Biosystems 7900HT - Page 174

DRAFT
September 1, 2004 11:39 am, CH_End-Point.fm
Chapter 5 Analyzing End-Point Data
5-16 Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User Guide
Scrutinizing the
Allele Calls
1. Verify the calls for the NTC, Allele X, and Allele Y controls.
a. In the plate grid, select the wells containing the No Template Control
samples.
The software highlights the datapoints within the allele plot.
b. Check that the datapoints cluster in the expected position on the plot.
c. If using positive controls, repeat steps a and b for the wells containing the
Allele X, and Allele Y controls.
2. Designate samples that did not cluster tightly as Undetermined.
Samples that did not cluster tightly may:
• Contain rare sequence variations
• Contain sequence duplications
• Contain contaminants
• Be the result of errors in pipetting or sample preparation
Allele X controls
should cluster here
Allele Y controls
should cluster here
No Template
Controls should
cluster here
Undetermined
samples