title: GSM4143073: 26SrRNA_eri1(mg366)_GFPNRDE3_E; Caenorhabditis elegans; OTHER
library_construction_protocol: total RNA were extracted from embryos or L3 larva, digested by DNase I, and then ligated to the following 3′ RNA linkers with T4 RNA ligase (Thermo #EL0021) (1 μg total RNA, 2 μl 3′ RNA linker (10 μM), 1 μl 10× T4 RNA ligation buffer, 2 μl T4 RNA ligase) by incubating at 37 °C for 30 min. Briefly, total RNA were extracted from embryos or L3 larva, digested by DNase I, and then ligated to the following 3′ RNA linkers with T4 RNA ligase (Thermo #EL0021) (1 μg total RNA, 2 μl 3′ RNA linker (10 μM), 1 μl 10× T4 RNA ligation buffer, 2 μl T4 RNA ligase) by incubating at 37 °C for 30 min. 3′ RNA linker: 5′-pGATCCACACTCGGGCACCAAGGATTTAACCGCGAATTCCAGC-NH2-3′ (the underlined sequence served as a barcode for sample labeling). The RNAs were reverse transcribed with the following primers: 3′ linker RT: 5′-GCTGGAATTCGCGGTTAAATCCTTGGTGCCCGAGTGTGGATC-3′. The cDNAs were PCR amplified with the primers 26S rRNA-F: 5′-CAGATCACTCTGGTTCAATGTC-3′ and 3′ linker RT primers, gel purified and then deep sequenced using an Illumina platform, according to the manufacturer′s instructions, by Novogene (Beijing, China). The number of reads with distinct 3′ -end modifications were counted by custom Perl scripts.
attributes.tag: GEO Accession
attributes.value: GSM4143073