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Forensic STR Profile Analysis Essentials: Panel Customization and Quality Size Call Verification

Panel files and size standards are pre-loaded in GeneMarker® HID software and include:

  • GlobalFiler™ 6-Dye™
  • Identifiler®
  • MiniFiler®
  • Y-Filer™ Plus
  • PowerPlex®16
  • PowerPlex®ESI
  • PowerPlex®Y-23
  • SureID® STR
  • Investigator®24Plex
  • Investigator®DIPplex
  • PowerPlex®Fusion 5 and 6 Dye
  • iPLEX STR™

Preloaded standards include (but are not limited to) GeneScan™500-GS600, Internal Lane Standard ILS500-ILS600 and BTO-550. Users can easily construct a panel for analysis with custom chemistries. GeneMarkerHID software also imports files from all ABI®PRISM, Applied Biosystems SeqStudio™, Promega Spectrum Compact CE System genetic analyzers, or a rapid system (such as RapidHIT™ and RapidHIT™ID) and .fsa, .hid, and .sg1 file formats.

 

Figures 1 and 2 below illustrate the filtering and flagging parameters for each locus. These rules can be customized in the panel editor, including Minimum Heterozygous and Homozygous RFUs for Analytical Range and Stochastic Ranges, as well as percentages for Heterozygous Imbalance and Stutter.

Figure 3 is an example of the size call quality confirmation linear plots for each file and quality statistics for disabled fragments, as is recommended for GeneScan™500LIZ®, and options to save or print the Sizing Quality Report. Additional Sizing Quality flagging may be implemented per the lab SOP to alert the analyst of missing size standard peaks, degradation of signal or migration variation outside of the linear range.

 

User interface to set marker parameters, flags and filters

Figure 1: Customize panels in the Panel Editor. Each marker can be edited for minimum calling RFU (analytical range), Inconclusive (stochastic range) Heterozygous Imbalance, and Stutter percentages.

 

Examples of Allelic ladder flagging

Figure 2: When an allele ladder sample migration is beyond the auto-panel adjust range the out of bin peaks are flagged with red allele labels and red vertical bars in Main Analysis Screen. The figure above from the Panel Editor shows that there may be problems with this capillary.

 

Size Quality Verification

Figure 3: Confirm Sizing Quality at a glance: Quality Scores, Electropherograms, Linear Plots and Quality Statistics for disabled fragment(s). Use this screen as a review tool or print/save it for inclusion with the project audit trail.

 

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