RNAseq S2 Clamp RNAi knockdown
Three biological replicates of RNA-seq were performed using S2 cells incubated for 72 hours with RNAi targeting Clamp or LacZ (control) transcripts. For each target, 10 µg of each purified dsRNA was combined to a total of 30 µg and directly added to 1.5 × 106 S2 cells seeded in a 6-well plate. Cells were incubated for 72 hours at 25 °C.  Total RNA was extracted from treated cells using the RNeasy Mini Kit (Qiagen, 74104). RNA-seq libraries were prepared from the isolated RNA using the NEBNext UltraExpress RNA Library Prep Kit (NEB, E3330S), following the manufacturer's protocol. Libraries were sequenced on an Illumina NextSeq 2000 platform, generating 122-base single-end reads.
Three biological replicates of RNA-seq were performed using S2 cells incubated for 72 hours with RNAi targeting Clamp or LacZ (control) transcripts. For each target, 10 µg of each purified dsRNA was combined to a total of 30 µg and directly added to 1.5 × 106 S2 cells seeded in a 6-well plate. Cells were incubated for 72 hours at 25 °C.  Total RNA was extracted from treated cells using the RNeasy Mini Kit (Qiagen, 74104). RNA-seq libraries were prepared from the isolated RNA using the NEBNext UltraExpress RNA Library Prep Kit (NEB, E3330S), following the manufacturer's protocol. Libraries were sequenced on an Illumina NextSeq 2000 platform, generating 122-base single-end reads.