Sample information curated by ChIP-Atlas

Antigen

Antigen Class
TFs and others
Antigen
Smad2

Cell type

Cell type Class
Pluripotent stem cell
Cell type
P19
Tissue
Embryo
Disease
Teratocarcinoma; Embryonal Carcinoma

Attributes by original data submitter

Sample

source_name
p19 cells
cell type
Embryonic teratoma
cell line
p19
signal inhibition pretreatment
10 μM SB-431542 overnight + washout
ligand treatment
1 hr 20 ng/ml Activin A
chip antibody
SMAD2 (Cell signaling 5339)

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Samples for ChIP-seq with the corresponding inputs were prepared by crosslinking in 1% formaldehyde for 10 minutes at RT, cytoplasmic lysis, sonication of nuclei to shear the DNA to 100-500 bp, followed by incubation with anti-SMAD2 antibody overnight. pre-blocked protein A dynabeads (invitrogen) were added for 4 hours, followed by washing, DNA elution, reversal of crosslinks and proteinase K digestion. Resulting enriched ChIP DNA was cleaned up using Qiagen PCR purification kit. Corresponding input chromatin was processed alongside at the same time. Following end repair, poly-A-tailing and adapter ligation, Illumina TruSeq ChIP sample preparation kits were used to generate the libraries. The Illumina kit Phusion enzyme was replaced by Kapa HiFi HotStart ready mix (Kapa Biosystems, Cape Town, South Africa). The PCR was run before gel isolation using the Invitrogen SizeSelect E-gel system (SizeSelect gel protocol, Thermo Fisher Scientific). Post PCR we used AMPure XP beads (AMPure bead protocol, Beckman Coulter, Inc.) at a 1:1 ratio to maintain size integrity. Samples were multiplexed and 51-bp single end reads were generated on an Illumina HiSeq 2500.

Sequencing Platform

instrument_model
Illumina HiSeq 2500

mm10

Number of total reads
78813534
Reads aligned (%)
87.0
Duplicates removed (%)
21.8
Number of peaks
1123 (qval < 1E-05)

mm9

Number of total reads
78813534
Reads aligned (%)
86.8
Duplicates removed (%)
21.7
Number of peaks
1185 (qval < 1E-05)

Base call quality data from DBCLS SRA