Library Preparation and Sequencing cDNA was synthesized using Takara SmartSeq v4 reagents from [1ng] of RNA. Full length cDNA was fragmented to a mean size of 200bp with a Covaris R230 ultrasonicator and Illumina sequencing libraries were prepared from 2ng of sheared cDNA using IDT DNAseq library prep reagents on a Beckman Coulter Biomek i7 according to manufacturer’s protocol. The finished dsDNA libraries were quantified by Qubit fluorometer and Agilent TapeStation 2200. Uniquely dual indexed libraries were pooled in equimolar ratios and evaluated for cluster efficiency and pool balance with shallow sequencing on an Illumina MiSeq. Final sequencing was performed on an Illumina NovaSeq X Plus with paired-end 150bp reads at the Dana-Farber Cancer Institute Molecular Biology Core Facilities.