title: GSM3335454: UCI_09_DLPFC; Homo sapiens; OTHER
library_construction_protocol: Approximately 50mg of pulverized brain tissue (grey matter only), or 50μl of frozen whole blood, was used for DNA extraction. Total DNA from brain tissue was extracted using the DNeasy Blood & Tissue Kit (Cat# 69504 or 69506, Qiagen), or the AllPrep DNA/RNA/Protein Kit (Qiagen, Valencia, CA, USA). For blood samples, DNA was isolated using 50μl of frozen whole blood (that had been stored in EDTA blood collection tubes) using the DNeasy Blood and Tissue Kit (Qiagen, Valencia, CA, USA). Long-range PCR protocol: Prior to NGS library preparation, the mtDNA was enriched for each sample using a 30-cycle long-range (LR) PCR that utilizes back-to-back primers that hybridize to the control region of the mitochondrial genome. Primer sequences used were 5'-CCGCACAAGAGTGCTACTCTCCTC-3' and 5'-GATATTGATTTCACGGAGGATGGTG-3' (Integrated DNA Technologies, Coralville, IA, USA), for the forward and reverse primers, respectively. PCR products then were purified using Agencourt AMPure XP beads (Beckman Coulter, Indianapolis, IN, USA). Fragmentation (Covaris) protocol: DNA shearing performed in S220 Covaris microTUBEs with the following settings: Duty Factor=5%, Peak Incident Power=175 W, Cycles per Burst=200, Time=35 s. Libraries were prepared using the TruSeq Nano DNA HT Library Preparation Kit (Illumina, San Diego, CA, USA) according to the manufacturer's instructions. Each library was barcoded so that samples could be multiplex sequenced together with 96 samples per lane.
attributes.tag: GEO Accession
attributes.value: GSM3335454