RNA-seq of mus musculus: sorted MAIT cells without BRD509 boost
Total RNA was subsequently submitted to Novogene following their sample preparation and shipping instructions. All further laboratory work was performed by Novogene using their commercial protocol. RNA was reverse transcribed and cDNA amplified by in vitro transcription with the TaKaRa SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing (TaKaRa, Cat. No. 634889) or a low input method using NEB Next Ultra RNA Library Prep Kit for Illumina (NEB, Cat. No. 7530). First-strand cDNA synthesis and tailing by reverse transcription were performed using SMART (Switching Mechanism at 5 End of RNA Template) technology. Following first-strand synthesis, cDNA was amplified by PCR to produce the library. Quality control of the library was performed by quantification with a Qubit 2.0 fluorimeter and by qPCR. Insert size was measured by the Agilent 2100 Bioanalyzer automated gel electrophoresis system. The library was sequenced using the NovaSeq platform with Illumina sequencing technology to generate 150bp paired-end reads.