GSM4143072
GSM4143072: WAGO_IP_small_RNA_cde1(tm936)_GFPWAGO1; Caenorhabditis elegans; ncRNA-Seq
Total small RNAs were isolated from young adult stage worms. Briefly, worms were harvested and dounced on ice with pestle B in TRIzol reagent (Invitrogen) followed by isopropanol precipitation. The precipitates were treated with calf intestinal alkaline phosphatase (CIAP, Invitrogen), re-extracted with TRIzol, and treated with T4 polynucleotide kinase (T4 PNK, New England Biolabs) in the presence of 1 mM ATP. Small RNAs were cloned and deep sequenced using an Illumina platform, according to the manufacturer's instructions, by  Novogene (Beijing, China). sRNAs ranging from 17 to 30 nt were gel-purified and ligated to a 3' adaptor (5'-pUCGUAUGCCGUCUUCUGCUUGidT-3'; p, phosphate; idT, inverted deoxythymidine) and a 5' adaptor (5'-GUUCAGAGUUCUACAGUCCGACGAUC-3'). The ligation products were gel-purified, reverse transcribed, and amplified using Illumina's sRNA primer set (5'-CAAGCAGAAGACG GCATACGA-3'; 5'-AATGATACGGCGACCACCGA-3'). The samples were sequenced using an Illumina Hiseq platform.
GEO
GSM4143072
GEO Accession
GSM4143072