Application Guide - QIAxcel Advanced

QIAxcel Advanced Application Guide 10/2016 113 Automated sample loading, processing and analysis prevent human errors that can occur with manual sample handling, thus ensuring reliability and reproducibility of the measurements. Since no hazardous compounds are handled manually, this analysis method affords both convenience and safety. Using the recommended method, up to 96 samples per run can be analyzed unattended in about 90 minutes, or 12 samples in as little as 13 minutes. Digital data collection and management of experiments ensure traceability and standardized results. Automated identification via peak calling QIAxcel ScreenGel software automatically identifies DNA fragments from unknown samples using its peak calling function (peak calling table). This enables accurate detection and identification of the amplicons present in each sample. The peak calling table contains the sizes of all 11 possible amplicons generated by multiplex PCR and the tolerance in % for automated peak detection (Table 5). The QIAxcel ScreenGel software compares the electrophoretic pattern obtained for each sample with the peak calling table and reports detected peaks in a Peak Calling Result Table that summarizes the results of the experiment (Table 6). This automated primary analysis of the sample makes results interpretation faster and more reliable for screening of a large number of samples. Peak Calling Instruction E.coli 11-multiplex PCR Name Position Tolerance Name Position Tolerance ehxA 168 bp 18.000 % wzxO145 523 bp 4.400 % wzxO111 230 bp 13.500 % wbqEO121 wbqFO121 587 bp 5.500 % rfbO157 296 bp 11.100 % stx1 655 bp 5.200 % eae 375 bp 5.600 % wzxO103 740 bp 5.700 % wzxO26 417 bp 5.000 % wzxO45 890 bp 8.400 % stx2 477 bp 4.800 % Table 5. Peak calling instructions for 11-multiplex PCR. RFU x 1E0 15 bp 667 bp 419 bp 169 bp 3000 bp RFU x 1E0 15 bp 53 bp 667 bp 288 bp 379 bp 419 bp 169 bp 3000 bp RFU x 1E0 15 bp 667 bp 419 bp 169 bp 3000 bp RFU x 1E0 15 bp 53 bp 667 bp 288 bp 379 bp 419 bp 169 bp 3000 bp Figure 4. A The threshold is used during peak detection. Signals that exceed the threshold value (dark blue line) are detected as peaks. B If non-relevant peaks are identified, the threshold can be increased (e.g., to 5 %) to exclude the non-relevant or unspecific data from the analysis.

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