title: GSM3325471: Wild type whole kidney rep4; Mus musculus; RNA-Seq
library_construction_protocol: RNA was isolated from FACS isolated cells or whole kidney using the Qiagen RNeasy micro (cells) or mini (whole kidney) kit following the manufacturer's instructions. Cells were resuspended and vortexed in buffer RLT and whole kidneys homogenized in buffer RLT using a motorized pestle. Whole kidneys were additionally centrifuged through a QIAshredder to shear genomic DNA and reduce viscosity. The lysates were centrifuged through the provided column to isolate RNA, and the columns subsequently washed, DNase treated, washed, and eluted with RNase-free H2O. Libraries were constructed from 20-50ng of total RNA by the USC Epigenome Center with the Illumina TruSeq RNA Library Prep Kit V2 following the manufacturer's instructions with following modifications: End-IT repair from Epicentre was used instead of TruSeq end repair (more favorable volumes). Half of adapter-ligation was amplified for a variable number of cycles depending on input amounts. KAPA Biosystems PCR Master Mix was utilized in place of Illumina's.
attributes.tag: GEO Accession
attributes.value: GSM3325471