Application Guide - QIAxcel Advanced

QIAxcel Advanced Application Guide 10/2016 61 Materials and methods PCR products produced using the A-B-DR Combi Tray Kit (Olerup SSP AB) were analyzed using the QIAxcel system with the QIAxcel DNA Fast Analysis Kit and the DM150 method. QX Alignment Marker 15 bp/3 kb and QX DNA Size Marker 50 bp – 1.5 kb were included in the analysis. Electrophoretic separation was performed using 10 kV voltage and 150 s separation time. Samples were injected with an injection time of 10 s at 10 kV. HLA types were determined after analysis using BioCalculator software and HLA genotyping software. Results Three different types of HLA typing kits are available — sequence-specific oligonucleotides (SSO), sequencing-based typing (SBT), and sequence-specific primers (SSP) — and these are used in combination with different detection systems. For gel electrophoresis, sequence-specific primer kits are used. These kits are primarily dedicated to analysis of the A, B, C, DR, DQ, and DP loci of polymorphic HLA genes. Figure 1 presents the results of analysis of 96 samples for one person. Separation of 12 samples was completed on the QIAxcel in 3 minutes using the QIAxcel DNA Fast Analysis Kit. Ready-to-run gel cartridges allow 96 samples to be processed automatically without manual intervention, thus reducing handling errors. The QIAxcel system minimizes hands-on time significantly compared to traditional methods. Electrophoresis and data acquisition are fully automated. Since results obtained using the QIAxcel system are compatible with HLA genotyping software, data can be imported and exported easily, and HLA genotypes are obtained within a few minutes. Since the QIAxcel capillary electrophoresis system uses only minute amounts of DNA through electrokinetic injection, the samples are retained for downstream procedures. Conclusions • The combination of the QIAxcel system and QIAxcel DNA Fast Analysis Kit is an optimal solution for fast PCR checking, enabling rapid, reproducible, and standardized analysis of multiple HLA loci. The QIAxcel system is, therefore, highly suited for HLA genotyping and examining the role of HLA genes in multiple diseases in clinical research. • BioCalculator software provides automated data generation and data acquisition, using formats that allow data exchange with other analysis software. • The QIAxcel system is a powerful and versatile tool for researching and determining HLA types. Analysis of PCR products using the QIAxcel system facilitates a better understanding of the role of highly polymorphic HLA genes in multiple diseases in clinical research.

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