title: RNA-Seq of CT26 tumor responder to anti-PD1 treatment
design: For RNA sequencing on mouse tissue, tumors were harvested in RNAlater (Qiagen) and cut into small pieces of 25-30 mg, then were immediately flash-frozen in liquid nitrogen and transferred to -80C for long storage. For RNA extraction, the small pieces of tissue were transferred to a 2ml microcentrifuge tube, containing 600l of RLT buffer (Qiagen) with -mercaptoethanol. Tissues were homogenized in the TissueLyser LT (speed 30 Hz, 3 min with 1 bead) and then transferred to a QIAshredder spin column (Qiagen).  RNA was extracted from the supernatant by using the RNeasy Mini kit (Qiagen), according to the manufacturer protocol and  fragmented.The sequencing library was prepared using Illumina TruSeq mRNA stranded kit. Sequencing was performed at target depth of 60 mln of paired-end reads.
library_name: RPD1-1 RNA-SEQ