Experiment information
Accession CRX062862
Organism Mus musculus
Title HFD_ko_miCLIP-seq_rep2
BioProject PRJCA001786
BioSample SAMC108821
Platform Illumina HiSeq X Ten
Library
Library name Construction protocol Strategy Source Selection Layout
mRNAs were purified using Dynabeads mRNA Purification Kit (Life Technologies, 61006) and fragmented to size around of 100 nt using the fragmentation reagent (Life Technologies, AM8740). 10 μg of purified mRNAs were mixed with 25 μg of anti-m6A antibody (Abcam, ab151230) in 450 μl immunoprecipitation buffer (50 mM Tris, pH 7.4, 100 mM NaCl, 0.05% NP-40) and incubated by rotating at 4 °C for 2 h. The solution was then transferred to a clear flat-bottom 96-well plate (Corning) on ice and irradiated three times with 0.15 J/cm2 at 254 nm in a CL-1000 Ultraviolet Crosslinker (UVP). The mixture was then immunoprecipitated by incubation with Dynabeads Protein A (Life Technologies, 1001D) at 4 °C for 2 h. After extensive washing and on-bead end-repair and linker ligation, the bound RNA fragments were eluted from the beads by proteinase K digestion at 55 °C for 1 h. RNAs were isolated by further phenol-chloroform extraction and ethanol precipitation. Purified RNAs were used to construct the library using SMARTer smRNA-Seq Kit for Illumina (Takara, 635029) according to the manufacturer’s instructions. Sequencing was carried out on Illumina HiSeq X-ten platform with paired-end 150 bp read length. RNA-Seq TRANSCRIPTOMIC PCR PAIRED
Processing Planned read length (bp) for mate 1: 150
Planned read length (bp) for mate 2: 150
Insert size (bp): 50
Release date2020-11-13
Run
Run accession Run data file information
File nameFile size (MB)
CRR073699 CRR073699_f1.gz
CRR073699_r2.gz
5,355.19
6,406.45
Submittersun baofa (sunbf@big.ac.cn)
OrganizationBeijing Institute of Genomics, Chinese Academy of Sciences
Date submitted2019-09-27