title: Bulk RNA-seq from Trex1 KO in heart
design: Bulk RNA-seq from Trex1 KO in heart
library_construction_protocol: Hearts were harvested from mice and collected into cold RPMI media. Following procedures previously described (J. Lim, R. Rodriguez, K. Williams, J. Silva, A. G. Gutierrez, P. Tyler, F. Baharom, T. Sun, E. Lin, S. Martin, B. D. Kayser, R. J. Johnston, I. Mellman, L. Delamarre, N. R. West, S. Müller, Y. Qu, K. Heger, The Exonuclease TREX1 Constitutes an Innate Immune Checkpoint Limiting cGAS/STING-Mediated Antitumor Immunity. Cancer Immunol. Res. 12, OF1–OF10 (2024).), hearts were homogenized with nuclease-free hard tissue homogenizing 2.8 mm ceramic beads (OMNI International) by using a Bead Mill Homogenizer (OMNI International, 19-628) in RLT buffer. Tissue homogenates were centrifuged at 13,000 rpm for 15 minutes at 4°C. The supernatants were collected and processed for RNA isolation with RNeasy Mini Kit (Qiagen). Total RNA was quantified with the Qubit RNA HS Assay Kit (Thermo Fisher Scientific) and quality was assessed using RNA ScreenTape on a TapeStation 4200 (Agilent Technologies). For sequencing library generation, the TruseqStranded mRNA Kit (Illumina) was used with an input of 100 to 1,000 ng of total RNA. Libraries were quantified with Qubit dsDNA HS Assay Kit (Thermo Fisher Scientific) and the average library size was determined using D1000 ScreenTape on a TapeStation 4200 (Agilent Technologies).