Bulk RNA-seq of mouse SL4 subcutaneous tumor ICB-treated 1
Sequencing of extracted RNA from bulk tumor tissue was performed by the Massachusetts Institute of Technology BioMicro Center. A Fragment Analyzer (Advanced Analytical Technologies Inc.) was used to confirm the quality of the RNA samples. The Kapa Hyperprep kit (Roche) was then used to prepare and index Illumina libraries from ~250ng of total RNA. The libraries were confirmed using the Fragment Analyzer, and qPCR was used to quantify the libraries. The libraries were then pooled and sequenced on Illumina NextSeq500 with 40+40 paired end reads. Custom scripts were used to demultiplex the samples and allow single mismatch to the index sequencing.