Antigen

Antigen Class

Unclassified

Antigen

Unclassified

Cell type

Cell type Class

Bone

Cell type

U2OS

Tissue

bone

Lineage

mesoderm

Description

osteosarcoma from the tibia of a 15 year old, J. Ponten and E. Saksela derived this line (originally 2T) in 1964 from a moderately differentiated sarcoma, viruses were not detected during co-cultivation with WI-38 cells or in CF tests against SV40, RSV or adenoviruses, mycoplasma contamination was detected and eliminated in 1972, (PMID: 6081590)

Sample

source_name

U2OS cell

cell line

U2OS

treatment

DMSO

chip-grade antibody

Anti-Bmal1 (Abcam, Cat #ab3350, Lot# GR241928-12)

2nd antibody

anti-rabbit IgG

Sequenced DNA Library

library_strategy

ChIP-Seq

library_source

GENOMIC

library_selection

ChIP

library_construction_protocol

Cells in 15 cm dish were sequentially cross-linked in 20 ml PBS buffer containing 1.5 mM EGS for 20 min, and followed by cross-linked in 20 ml PBS buffer containing 1% formaldehyde for 15 min at room temperature. The cross-link was stopped by adding glycine to a final concentration of 125 mM and incubate at room temperature for 5 min. ChIP assay and DNA sample preparation were performed strictly following the ChIP-IT High Sensitivity Kit (Active Motif). Eluted DNA was analyzed with high-throughput sequencings. Libraries were prepared according to Illumina's instructions.

Sequencing Platform

instrument_model

Illumina HiSeq 2500

hg38

Number of total reads

23686397

Reads aligned (%)

92.5

Duplicates removed (%)

39.1

Number of peaks

19527 (qval < 1E-05)

hg19

Number of total reads

23686397

Reads aligned (%)

92.3

Duplicates removed (%)

39.5

Number of peaks

19506 (qval < 1E-05)