title: GSM4061885: ETH16h2.mRNA.m6A; Petunia x hybrida; OTHER
library_construction_protocol: The total RNA of the samples was isolated using the Trizol Kit (Promega, USA) and was treated with RNase-free DNase I (Takara Bio, Japan) for 30 min at 37℃ to remove residual DNA. Briefly, fragmented RNA was incubated with anti-m1A polyclonal antibody (Synaptic Systems, 202003) in IPP buffer for 2 hours at 4°C. The mixture was then immunoprecipitated by incubation with protein-A beads (Thermo Fisher) at 4°C for an additional 2 hours. Then, bound RNA was eluted from the beads with N1-methyladenosine (BERRY & ASSOCIATES, PR3732) in IPP buffer and then extracted with Trizol reagent (Thermo Fisher) by following the manufacturer's instruction. Purified RNA was used for RNA-seq library generation with NEBNext® Ultra™ RNA Library Prep Kit (NEB). Both the input sample without immunoprecipitation and the m1A IP samples were subjected to 150 bp paired-end sequencing on Illumina HiSeq sequencer. library construction protocol
identifiers.namespace: GEO
identifiers.id: GSM4061885