QIAamp DSP DNA Blood Mini Kit

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Publication protocol

Library preparation for plasma DNA: The obtained plasma samples were processed for DNA isolation using an optimization of a previously published standard method24. In the standard method: DNA was extracted by DNeasy Blood and Tissue Kit (No. 69504; Qiagen), and indexed DNA libraries were prepared as described in15, using the following steps: i) End repair. ii) Adenine tailing. iii) Adaptor ligation. iv) Library amplification. DNA was purified by 1.5X SPRI beads after each step. For our optimized method: DNA was extracted by QIAamp DSP DNA Blood Mini Kit (No. 61104; Qiagen). DNA libraries were prepared by KAPA Hyper Prep Kits (No. KK8502; Kapa), using the following steps: i) End repair and adenine tailing combined in a single step. ii) Proceeded directly to adaptor ligation. iii) Adaptor ligated DNA was purified by 1.5X SPRI beads and amplified by indexed primers. For size selection to enrich mtDNA fragments in the library, the library was size selected from 150 to 190 bp by Pippin electrophoresis (Sage Sciences Blue Pippen). Libraries were sequenced by the Illumina Hiseq platform with paired-end read lengths of 150 bp.

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