title: GSM3317312: Bone marrow sinusoidal endothelial cells, DT treated, biological rep 2; Mus musculus; RNA-Seq
library_construction_protocol: Sinusoidal endothelial cells were sorted using flow cytometry, gated as Lineage- (CD45, Ter119, Gr1, CD11b) CD31+ Sca1- cells, and the RNA from these cells was extracted using RNeasy micro Kit (74004, Qiagen). Library preparation was performed with 1ng of total RNA, integrity was determined using an Agilent bioanalyzer. ds-cDNA was prepared using the SMARTer Ultra Low RNA kit for Illumina Sequencing (Clontech) per manufacturer's protocol. cDNA was fragmented using a Covaris E220 sonicator using peak incident power 18, duty factor 20%, cycles/burst 50, time 120 seconds. cDNA was blunt ended, had an A base added to the 3' ends, and then had Illumina sequencing adapters ligated to the ends. Ligated fragments were then amplified for 14 cycles using primers incorporating unique index tags. Fragments were sequenced on an Illumina HiSeq-3000 using single reads extending 50 bases.
attributes.tag: GEO Accession
attributes.value: GSM3317312