GSM5916190: Whole_MeRIP_Input_BD10-2; Drosophila melanogaster; RIP-Seq
Total RNAs were isolated and dissolved in RNase-free water after the bulk samples had been homogenized in 0.5–1 mL TRIzol AG RNAex Pro Reagent (#AG21102, Accurate Bioeng. Co., Ltd., Hunan, China) with glass beads using a homogenizer. Extracted total RNAs were subjected to poly-A selection utilizing VAHTS® mRNA Capture Beads (#N401-02, Vazyme Biotech. Co., Ltd., Jiangsu, China). These purified poly-A RNAs were fragmented as ~100 nt fragments for the preparation of both m6A IP and untreated Input control sequencing libraries of MeRIP-seq. VAHTS® Universal V6 RNA-seq Library Prep Kit for Illumina (#NR604-02, Vazyme Biotech. Co., Ltd.) and VAHTS® RNA Adapters set6 for Illumina (#N812, Vazyme Biotech. Co., Ltd.) provided necessary reagents for the fragmentation of poly-A RNAs, first- and second-strand cDNA synthesis, adaptor ligation, and library amplification with index sequences. For the m6A IP sequencing libraries, we carried out m6A IP with poly-A RNA fragments with EpiMark® N6-Methyladenosine Enrichment Kit (#E1610S, New England Biolabs, MA, USA). Then the m6A-enriched poly-A RNA fragments and un-enriched poly-A RNA fragments were employed for the construction of strand-specific IP and Input libraries of MeRIP-seq, respectively.