Application Guide - QIAxcel Advanced

QIAxcel Advanced Application Guide 10/2016 57 PCR products were analyzed using the QIAxcel system with the QIAxcel DNA Screening Kit and the QX Alignment Marker 15 bp/450 bp. Electrophoretic separation was performed using 2 kV separation voltage and 700 second separation time. Alignment marker and samples were injected with an injection time of 35 seconds at 2 kV and 3 kV, respectively. Results and discussion Results of the analysis of 5 samples (S1–S5) are shown in Figure 1. The presence and size of fragments was accurately determined using BioCalculator Software. B-cell monoclonality was observed for the S1, S4, and S5 samples. B-cell oligoclonality was detected for the S3 sample. For sample S5a and S5b, taken at different times during the course of the disease, the same monoclonal rearrangements were observed for the IGH and IGK genes. (Table 1). Figure 1. Identification of gene rearrangements in proliferating B-cells. DNA from lymphoid tissues was amplified using primers developed in the BIOMED-2 study and analyzed using the QIAxcel system with the QIAxcel DNA Screening Kit. M: size marker; C–: negative control; C+: positive control. S1–S5b: duplicate samples of multiplex PCR products. Primers were chosen to detect rearrangements in FR3 (A) and FR2 (B) VD FR regions of the IGH gene as well as the IGK-A (C) and IGK-B (D) genes. M M M M C– C– C– C– C+ C+ C+ C+ S1 S1 S1 S1 S2 S2 S2 S2 S3 S3 S3 S3 S4 S4 S4 S4 S5a S5a S5a S5a S5b S5b S5b S5b IGH-FR3 IGH-FR2 IGK-A IGK-B A B C D

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