Sample information curated by ChIP-Atlas

Antigen

Antigen Class
Unclassified
Antigen
Unclassified

Cell type

Cell type Class
Pluripotent stem cell
Cell type
ES cells
NA
NA

Attributes by original data submitter

Sample

source_name
E14TG2a
cell line
D395A
treatment
untreated
protocol
ChIP

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
For RNA-seq, RNA was isolated using TRNzol reagent. After DNase I treatment, rRNA was removed using QIAseq FastSelect RNA Removal Kit (QIAGEN). For RIP-seq, chromatin RNA was extracted, then fragmented to about 100-300 nt-long, and incubated with anti-m6A antibody (Millipore). Then the mixture was subjected to Dynabeads protein G binding. After washing, the bound RNA was eluted by RLT and retrieved by RNeasy Plus Micro Kit (Qiagen). H3K9me2 ChIP-seq was performed using the iDeal ChIP-seq Kit for Histones (Diagenode). For KDM3B, METTL3 and Pol II ChIP-seq, cells were crosslinked, sonicated, and subjected to ChIP. RNA-seq libraries were generated using NEBNext Ultra Directional RNA Library Prep Kit for Illumina (NEB). RIP-seq libraries were generated using SMARTer Stranded Total RNA-Seq Kit v2 -Pico Input Mammalian (Takara) following the manual. ChIP-seq libraries were generated using NEBNext Ultra DNA Library Prep Kit for Illumina. RNA-seq, RIP-seq and ChIP-seq

Sequencing Platform

instrument_model
Illumina NovaSeq 6000

mm10

Number of total reads
50740467
Reads aligned (%)
95.2
Duplicates removed (%)
21.4
Number of peaks
456 (qval < 1E-05)

mm9

Number of total reads
50740467
Reads aligned (%)
95.2
Duplicates removed (%)
21.4
Number of peaks
431 (qval < 1E-05)

Base call quality data from DBCLS SRA