Sample information curated by ChIP-Atlas

Antigen

Antigen Class
Histone
Antigen
H3K4me3

Cell type

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

Attributes by original data submitter

Sample

source_name
embryonic stem cell
cell type
embryonic stem cell
cell line
v6.5
genotype/variation
Set1A SET deletion
chip antibody
H3K4me3 (Homemade, #519)

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
RNA was extracted using RNeasy mini kit (Qiagen) following manufacturer's instructions. For ChIP, ESCs were fixed in 1% formaldehyde, followed by quenching and cell lysis before chromatin sonication using a E220 focused-ultrasonicator (Covaris). Sheared chromatin was then subjected to immunoprecipitation, and immunoprecipitated DNA was then washed, eluted, reverse cross-linked, and purified prior to submission for library preparation. For EB ChIP experiments, EBs were dounce homogenized prior to sonication. ChIP-seq libraries were prepared using the KAPA HTP Library Preparation Kit complemented with NEXTflex DNA Barcodes from Bioo Scientific. 10 ng of DNA was used as starting material for input and IP samples. Libraries were amplified using 13 cycles on the thermocycler. Post amplification libraries were size selected at 250- 450bp in length using Agencourt AMPure XP beads from Beckman Coulter. Libraries were validated using the Agilent High Sensitivity DNA Kit. RNA-seq libraries were prepared using the Illumina TruSeq Stranded Total RNA Preparation Kit with Ribo-Depletion. Input RNA quality was validated using the Agilent RNA 6000 Nano Kit. 200ng-1 µg of total RNA was used as starting material. Libraries were validated using the Agilent DNA 1000 Kit.

Sequencing Platform

instrument_model
NextSeq 500

mm10

Number of total reads
49408435
Reads aligned (%)
96.2
Duplicates removed (%)
21.7
Number of peaks
17230 (qval < 1E-05)

mm9

Number of total reads
49408435
Reads aligned (%)
95.8
Duplicates removed (%)
21.6
Number of peaks
17309 (qval < 1E-05)

Base call quality data from DBCLS SRA