GSM9395684: WT, 5B2, 5529; Homo sapiens; RNA-Seq
At necropsy, tumors were weighed before partitioning for RNA isolation or histological processing. RNA was isolated from flash frozen samples using Qiagen RNeasy Mini Kit (Qiagen 74104). For preparation of sequencing libraries, 500ng of RNA isolated from flash frozen tumors was used with the NEBNext Ultra II Directional RNA Library Prep kit for Illumina along with NEBNext Poly(A) mRNA Magnetic Isolation Module according to the manufacturers instructions. Library quality and quantity were assessed using RNA ScreenTape Assay (Agilent) and Agilent 4200 TapeStation System following the manufacturers protocol. 3 nM equimolar pooled libraries were sequence on the Illumina NextSeq 2000 system (Illumina) using NextSeq 1000/2000 P2 Reagent Cartridge 100 cycles and P2 Flow Cell (Illumina).