title: GSM7883450: Midgut, resistant, rep3, IP; Plutella xylostella; RIP-Seq
library_construction_protocol: Total RNA was isolated using TRIzol reagent (Invitrogen), and Poly (A) RNA was extracted from 50 μg total RNA with Dynabeads Oligo (dT) 25-61005 (Thermo Fisher Scientific). The poly (A) RNA was fragmented using the NEBNext Magnesium RNA Fragmentation Module (New England Biolabs). Then, the fragmented RNAs were incubated with the m6A-specific antibody (Synaptic Systems). Then, the precipitated RNA was reverse-transcribed to generate cDNA using the SuperScript II Reverse Transcriptase (Invitrogen), and the obtained cDNA was used to synthesize U-labeled second-stranded DNAs with Escherichia coli DNA, RNase H (New England Biolabs), dUTP Solution (Thermo Fisher Scientific), and polymerase I (New England Biolabs). The indexed adapters were prepared using the blunt ends with an A-base. The A-tailed fragmented DNA was ligated to the adapter with a T-base, and the AMPureXP beads were used to select the size of index adapters. The U-labeled second-stranded DNAs were treated with heat-labile UDG enzyme (New England Biolabs), the ligated fragments were amplified using PCR