Application Guide - QIAxcel Advanced

6 QIAxcel Advanced Application Guide 10/2016 Quality control of genomic DNA using the QIAxcel® system André Schaller, Christopher Jackson, Division of Human Genetics, Inselspital Bern, Berne, Switzerland Thomas Kretschmann, QIAGEN GmbH, Hilden, Germany This application note describes a rapid, reliable, and effective method for quality control of purified genomic DNA using the QIAxcel system. The results are highly reproducible and clearly show whether degradation products are present in a sample. Introduction The results of numerous molecular screening and assay methods rely on the quality of the genomic DNA (gDNA) that is used. Effective techniques for purification are crucial to secure gDNA that will give the best results in downstream procedures, but optimized quality control of purified gDNA is equally important. It helps to avoid time and money being wasted, particularly in the case of costly procedures, such as Next Generation Sequencing (1), where gDNA is fragmented and used for library preparation. The sizes of purified gDNA fragments depend on the purification method. They fall between 20 and 30 kb when spin columns with silica-based membranes are used, and can be up to 100 kb or more when salting-out precipitation is applied. The silica-based method is the most commonly used, and it can be done manually or automated with a sample purification instrument such as the QIAcube® . Using the QIAxcel system, quality control is possible for all gDNA that has been purified with silica membrane-based methods. It offers a straightforward and effective means for checking the quality of all samples purified with kits that use this approach (e.g., QIAamp® DNA Blood Kit, QIAamp DNA FFPE Tissue kit, QIAamp MinElute Virus Kits). Materials and Methods Genomic DNA was isolated from blood samples using QIAGEN’s QIAamp DNA Blood Mini Kit, which is based on silica membrane DNA purification. The protocols were automated and run on the QIAcube. The DNA samples were quantified with a NanoDrop® spectrophotometer. Concentrations of 10– 100 ng/μl were chosen for testing. Samples were analyzed on 1% agarose gels in 30 mM TAE buffer for 2.5 h at 80 V. The same samples were analyzed using the QIAxcel system

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