title: GSM1477264: Th9+TLA1 rep2; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was isolated from T cells using RNeasy kits (Qiagen, Valencia, CA) according to the manufacturer’s protocol Clontech’s SMARTer® Ultra™ Low Input RNA Kit for Sequencing v3 was employed to generate double-stranded cDNA libraries from 1.5 to 2.5 ng total RNA from each sample per manufacturer’s recommendations. Double-stranded cDNA libraries were individually fragmented and ligated with Ion Torrent Ion Xpress™ Barcode Adapters using Ion Xpress™ Plus Fragment Library Kit with limited modifications including size-selected final libraries with a double bead clean-up and ½ volume of Ion Adapters.  RNA-Seq libraries were assessed for concentration and length using Invitrogen’s Qubit® dsDNA HS Assay Kit (Carlsbad, CA) and Agilent High Sensitivity DNA Kit respectively (Carlsbad, CA). Samples were multiplexed to obtain >10 million (11-15 million) reads each for sequencing. The pooled libraries were amplified onto Ion Sphere™ and sequenced using the Ion PI™ Sequencing 200 Kit v2, Ion Torrent™.