title: GSM4658494: Human patients 1-3, hashed and pooled; Homo sapiens; RNA-Seq
library_construction_protocol: For mouse and human adhesion fibroblasts, fibroblasts were isolated from fresh specimens using FACS. RNA extraction was performed using Qiagen miRNeasy kit (cat. 1071023) with on column DAnase treatment per the manufacturer's recommendations. The Clontech Smarter Ultra Low Input RNA kit (Takara Bio, Cat. 634848) was used to generate cDNA from 150 pg total RNA following the manufacturer's recommendations.  Amplified cDNA was purified using SPRI Ampure Beads (Beckman Coulter, Cat. A63880) and the quality and quantity was measured using a High Sensitivity DNA chip on the Agilent 2100 Bioanalyzer (Agilent Technologies). cDNA was sheared to an average length of 300 basepairs using a Covaris S2 ultrasonicator (Covaris) and libraries were generated with the Clontech Low Input Library Prep kit (Takara Bio, Cat. 634947). The samples were uniquely barcoded, pooled, and sequenced on a single lane of the NextSeq 500 (Illumina). A total of 300 million paired-end, 151 base pair reads were obtained, resulting in 50 million reads per sample.
attributes.tag: GEO Accession
attributes.value: GSM4658494