EZ1 DSP DNA Blood Kit (48)

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Publication protocol

Genomic DNA (gDNA) was isolated from 350 µL EDTA patient blood using QIAGEN EZ1 DSP DNA Blood kit (QIAGEN) on QIAGEN EZ1 Advanced XL system.6 The extracted gDNA sample was quantified with Quant‐iT™ dsDNA PicoGreen™ Assay Kit (Thermo Fisher) using SpectraMax® Gemini EM Microplate Spectrofluorometer (Molecular Devices). The integrity of gDNA was confirmed by running 0.8% E‐gel (Thermo Fisher). A DNA library construction started with fragmentation using Covaris Sonicator E220 (Covaris) to shear 1 µg gDNA to the fragments size of 300‐500 base pairs (bp). A fragmented DNA sample was then treated with Illumina TruSeq®DNA PCR‐Free Library Prep Kits in order to yield a final library, in which fragments were ligated with a molecular barcoded adapter. The ligated fragment size and concentration of a DNA library sample was measured using Fragment Analyzer™ (Advanced Analytical) and KAPA Library Quantification Kits on Roche LightCycler® 480 Instrument (Roche). A library with the concentration more than 3 nmol/L (300‐600 bp) was passed for sequencing on Novaseq 6000 S2 Flow Cell (FC).

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