Sample information curated by ChIP-Atlas

Antigen

Antigen Class
RNA polymerase
Antigen
RNA polymerase II

Cell type

Cell type Class
Blood
Cell type
RAW 264.7
Primary Tissue
Blood
Tissue Diagnosis
Leukemia

Attributes by original data submitter

Sample

source_name
Raw 264.7
strain
Adult strain BALB/c male
cell line
Raw 264.7
cell type
macrophage; Abelson murine leukemia virus transformed
antibody
Pol II antibody from Santa Cruz (sc-899)
treatment
No treatment

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Roughly 80 x 10^6 (for Fli1, Elf1 and Pol II ChIP-seq), 40 x 10^6 (for GABP-α, Elf4 ChIP-seq) or 5 x 10^6 (for PU.1 and histone modifications ChIP-seq) cells were fixed in 1% formaldehyde for 10min and were lysed with RIPA buffer, after chromatin shearing by sonication. Lysate was immunoprecipitated with 5-10 μg of antibody. Antibodies were pre-bound overnight to 50-100 μl of G protein-coupled paramagnetic beads in PBS/BSA 0.5%. Beads were then added to lysates (the preclearing step was omitted) and incubation was let to proceed overnight. Beads were washed 6 times in a modified RIPA buffer (50mM Hepes pH 7.6, 500mM LiCl, 1mM EDTA, 1% NP-40, 0.7% Na-deoxycholate) and once in TE containing 50mM NaCl. DNA was eluted in TE containing 2% SDS and crosslinks reversed by incubation overnight at 65ºC. DNA was then purified by Qiaquick columns (Qiagen) and quantified using PicoGreen (Invitrogen). ChIP DNA libraries were prepared for HiSeq 2000 Illumina sequencing using a standard protocol (Garber et al., 2012) with slight modifications (Ostuni et al., 2013).

Sequencing Platform

instrument_model
Illumina HiSeq 2000

mm10

Number of total reads
42249813
Reads aligned (%)
94.3
Duplicates removed (%)
18.0
Number of peaks
20717 (qval < 1E-05)

mm9

Number of total reads
42249813
Reads aligned (%)
94.0
Duplicates removed (%)
18.0
Number of peaks
20954 (qval < 1E-05)

Base call quality data from DBCLS SRA