GSM4293944
GSM4293944: WT D0 rep2; Mus musculus; OTHER
Isolate mice liver and remove gall bladder. Cut the liver into small pieces and place them in RNAlater (Thermo). Liver tissues were ground in liquid nitrogen after added lysis buffer. Then extract the RNAs  with mirVana™ miRNA Isolation Kit (Thermo, AM1561). Isolate 14-40 nt RNAs by PAGE. Total RNA samples were first pretreated as following to remove some RNA modifications 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 efficient 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 amplification; 5) size selection of ~134-160bp PCR amplified fragments (corresponding to ~14-40nt small RNAs). The completed libraries were quantified by Agilent 2100 Bioanalyzer. Libraries were mixed in equal amounts according to the quantification results, and used for sequencing on the instrument. tRF-seq
GEO
GSM4293944
GEO Accession
GSM4293944