title: GSM6873892: Naïve CD4 T cell, KO_S1PR1_H; Mus musculus; RNA-Seq
library_construction_protocol: RNA was isolated from FACS-sorted naïve CD4+ T cells using TRIzol LS (Invitrogen) followed by DNase I (QIAGEN) treatment and cleanup with an Rneasy Plus Micro kit (QIAGEN), and quantified on a 2100 BioAnalyzer instrument (Agilent technologies, Inc.). The RNASeq library prep and sequencing were performed at the NYU Langone Genome Technology Center. 5-20 ngs of total RNA was used to prepare libraries using Trio RNA-Seq library prep kit (Tecan Genomics, Inc.) following manufacturer's instructions. The protocol contains the following steps: DNase treatment to remove any DNA in the sample, first strand and second stand cDNA synthesis from the input RNA, single primer isothermal amplification (SPIA) of the resultant cDNAs, enzymatic fragmentation and construction of unique barcoded libraries, PCR library amplification (4 cycles were used) and an AnyDeplete step to remove rRNA transcripts. The Agencourt AMPure XP bead (Beckman Coulter) purified libraries were quantified using by qPCR and the size distribution was checked using Agilent TapeStation 2200 system.