title: GSM3638384: CNS TH17 ATAC-seq rep1; Mus musculus; ATAC-seq
library_construction_protocol: Naive CD4+ T cells from control and miR-21-/- mice were differentiated under pathogenic TH17 (23) differentiation condition. Total RNA was prepared from these cells using Trizol reagent (Invitrogen). RNA-seq libraries were prepared using a TruSeq Stranded Total RNA Library Prep Kit (Illumina). For GFP+ TH17 cells sorted from in vivo suspensions, 50,000 cells were resuspended in 700 ul Trizol reagent, total RNA was prepared by miRNeasy Micro Kit (Qiagen), RNA-seq libraries were prepared using SMART-Seq v4 Ultra Low Input RNA Kit (clontech). Sequencing was performed on an Illumina HiSeq X Ten System in a 150 bp/150 bp Paired end mode. ATAC-seq library preparations were performed as described(Buenrostro et al., 2013). In brief, 50,000 cells were washed in cold PBS and lysed. Transposition was performed at 37 °C for 30 min. After purification of the DNA with the MinElute PCR purification kit (Qiagen), DNA was amplified for 5 cycles. Additional PCR cycles were evaluated by real time PCR. Final product was cleaned by Ampure Beads at a 1.5× ratio. Libraries were sequenced on a HiSeq X Ten System in a 150 bp/150 bp Paired end mode. RNA libraries were prepared for sequencing using standard Illumina protocols
attributes.tag: GEO Accession
attributes.value: GSM3638384