GSM9023419: TERRA_captured_Nanopore_direct_RNA-seq 2; Homo sapiens; RNA-Seq
Total RNA was extracted from 1 x 10^7 cells using TRIzol™ Reagent (Invitrogen™, Cat:15596018) following the manufacturer's protocol. For Nanopore sequencing, TERRA-enriched RNA was polyadenylated using E. coli Poly(A) Polymerase (New England Biolabs, #M0276S) at a final concentration of 0.25 U/μl in 1× Poly(A) Polymerase Reaction Buffer containing 1 mM dATP and 2 U/μl RNaseOUT (Thermo Fisher Scientific, Cat: 10777019), and incubated at 37°C for 30 minutes. The reaction was terminated by adding 10 mM EDTA. The RNA sample was then purified using AMPure XP RNA beads (Beckman Coulter, A66514) and subsequently processed using the Oxford Nanopore direct RNA sequencing protocol (Cat# SQK-RNA004). Then subjected to FLO-MIN004RA flow cell for sequencing.