title: GSM8370489: Con1; Rattus norvegicus; ncRNA-Seq
library_construction_protocol: RNA was harvested using Rneasy mini plus kit (Qiagen). 2 ug of total RNA was used for the construction of sequencing libraries. Agarose electrophoresis was used to check the integrality of total RNA samples, and then the samples were quanti ed on the NanoDrop ND-1000 instrument. Total RNA samples were  rst pretreated as following to remove some RNA modi cations that interfere with small RNA-seq library construction: 3  -aminoacyl (charged) deacylation to 3 -OH for 3 adaptor ligation, 3 -cP (2 ,3 -cyclic phosphate) removal to 3 -OH for 3 adaptor ligation, 5 -OH (hydroxyl group) phosphorylation to 5 -P for 5  -adaptor ligation, m1A and m3C demethylation for e cient reverse transcription. Pretreated total RNA of each sample was taken for tRF & tiRNA-seq library preparation. Library preparation procedures included: 1) 3 -adapter ligation; 2) 5 -adapter ligation; 3) cDNA synthesis; 4) PCR ampli cation; 5) size selection of ∼134-160bp PCR ampli ed fragments (corresponding to ∼14-40nt small RNAs). The completed libraries were quanti ed by Agilent 2100 Bioanalyzer. Libraries were mixed in equal amounts according to the quanti cation results, and used for sequencing on the instrument. small RNA sequencing