title: GSM8596788: Whole blood, RNAseq, T2D, subj8; Homo sapiens; RNA-Seq
library_construction_protocol: Whole blood samples were retrieved from cryostorage and thawed at room temperature. The total RNA was isolated from 300 μL of whole blood using a TRIzol reagent (Invitrogen, Carlsbad, CA, USA) with chloroform, purificated with PureLink Total RNA Blood Kit (Invitrogen™, Carlsbad, CA, USA) following the manufacturer's instructions, and eluted with 45 μL RNase-free water. Total RNA was tested for purity using the NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA) and quantified with Qubit™ RNA High Sensitivity Assay Kit and Qubit 2.0 Fluorometer (both Invitrogen™, Carlsbad, CA, USA). Libraries were constructed from 150 ng of previously extracted total RNA using TruSeq Stranded Total RNA library preparation kit (Illumina, Inc., San Diego, California) according to the manufacturer's protocol. Briefly, after ribosomal RNA depletion and RNA fragmentation, first and second strand cDNA were synthesized, and 3' ends were adenylated, followed by adapter ligation and enrichment of DNA fragments. The library's size distribution was analyzed with Agilent 2200 TapeStation and Agilent High Sensitivity D1000 ScreenTape (both Agilent Technologies, Inc., Waltham, MA, USA).