GSM9065477: cells, FOLFOXIRI treatment1; Homo sapiens; RNA-Seq
Total RNA was isolated from CRC cells in xenograft tumor samples using TRIzol reagent (Thermo Fisher Scientific), following the manufacturer's instructions. RNA integrity and purity were assessed using the Bioanalyzer 2100 system and RNA 6000 Nano LabChip kit (Agilent Technologies), and samples with RNA integrity numbers (RIN) greater than 7.0 were selected for library construction. First-strand complementary DNA (cDNA) was synthesized using SuperScript II Reverse Transcriptase (Invitrogen), followed by the synthesis of second-strand cDNA using Escherichia coli DNA polymerase I (New England Biolabs), RNase H (New England Biolabs), and a dUTP solution (Thermo Fisher Scientific). The blunt-ended cDNA fragments were subsequently adenylated at the 3′-ends to facilitate ligation with dual-index adapters containing T-overhangs. Size selection of adapter-ligated cDNA (~300 ± 50 bp) was performed using AMPure XP beads (Beckman Coulter), followed by digestion of U-labeled second strands with heat-labile UDG enzyme (New England Biolabs). Libraries were PCR-amplified (95℃ for 3 minutes, followed by 8 cycles of 98℃ for 15 s, 60℃ for 15 s, and 72℃ for 30 s, and a final extension at 72℃ for 5 minutes).
GEO
GSM9065477_r1