title: GSM4106639: iMAC-Moi; Homo sapiens; RNA-Seq
library_construction_protocol: RNA was extracted using the RNeasy Plus Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer's protocol. Briefly, cells were homogenized and transferred directly into a spin column. Tubes were centrifuged 2 min at maximum speed. Homogenized lysate were then transfered into spin column and centrifuged for 30min at 8000g, and save the flow-through and add 1 volume of 70% EtOH. Transfer 700 ul of sample into RNeasy spin column and centrifuged for 15s at 8000g. After 15s, 700 ul RPE buffer was added to spin column and centrifuged 15s at 8000g. Next, 500 ul RPE buffer was added to spin column and centrifuged 2min at 8000g. After 2min, transfer spin column in a new 1.5 ml collection tube and added 50ul RNase free water directly to the membrane and centrifuged for 1min at 8000g. The library was prepared using Truseq stranded mRNA prep kit (Illumina, San Diego, CA, USA) and run on a HiSeq2500 sequencer (Illumina) with a 101-bp paired-end read length. Raw data were processed using STAR v.2.6 to align genome reads against human Reference genome (hg38) and differentially expressed genes were identified using DESeq21 after raw count data were normalized using variance stabilizing transformation (VST) method. Multidimensional scaling (MDS) and clustering analysis were calculated based on Poisson model. Spearman correlation coefficients between pairs of samples were computed.
attributes.tag: GEO Accession
attributes.value: GSM4106639