Sample information curated by ChIP-Atlas

Antigen

Antigen Class
Unclassified
Antigen
Unclassified

Cell type

Cell type Class
Blood
Cell type
RAJI
Primary Tissue
Blood
Tissue Diagnosis
Malignant Lymphoma - Burkitts Type

Attributes by original data submitter

Sample

source_name
Raji B-cell line
cell line
Raji B-cell line
antibody
1F5
library strategy
ChIP-seq

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Raji cells were crosslinked at room temperature by adding 1/10th volume of crosslinking solution (11% formaldehyde, 100 mM NaCl, 1 mM EDTA pH 8.0, 0.5 mM EGTA pH 8.0, 50 mM Hepes pH 7.8) to the cell culture medium for 10 min. The reaction was stopped by the addition of 250 mM glycine for 5 min. Afterwards the cells were washed twice with PBS. 5 x 107 cells were lysed in 2,5 ml LB1 (50 mM Hepes, pH 7.5, 140 mM NaCl, 1 mM EDTA, pH 8.0, 10% Glycerol, 1% NP-40, 0.5% Triton X-100, 1 x phosphatase inhibitors, 1 x EDTA‐free protease inhibitors (Roche)) for 20 min at 4°C on a shaker. The cells were spun down (5 min, 4°C, 1350 g) and washed with LB2 (200 mM NaCl, 1 mM EDTA pH 8.0, 0.5 mM EGTA pH 8.0, 10 mM Tris pH 8.0, 1 x phosphatase inhibitors, 1 x EDTA‐free protease inhibitors) for 20 min at 4°C. The samples were spun down again and suspended in 1.5 ml LB3 (1 mM EDTA pH 8.0, 0.5 mM EGTA pH 8.0, 10 mM Tris pH 8.0, 100 mM NaCl, 0.1 % Na-Deoxycholate, 0.5% N-lauroylsarcosine, 1 x phosphatase inhibitors, 1 x EDTA‐free protease inhibitors). The samples were sonicated with a Misonix 4000 (Misonix Inc.) sonicator for 14 cycles (30 sec on, 30 sec off, amplitude 40). Triton X-100 was added to a final concentration of 1% and the chromatin was collected by centrifugation (20 000 g, 20 min, 4°C). An aliquot of 50 µl was taken as input control. The input was mixed with an equal volume of 2 x elution buffer (100 mM Tris pH 8.0, 20 mM EDTA pH 8.0, 2 % SDS) and incubated at 65°C for 13-15 h. The sample was diluted with same volume of TE (10 mM Tris pH 8.0, 1 mM EDTA pH 8.0) and digested by RNase A (0.2 µg/ml, 2 h, 37°C) and Proteinase K (0.2 µg/ml, 2 h, 55°C). DNA was purified by two phenol:chloroform:isoamylalcohol (25:24:1, pH 8.0) extractions steps and a subsequent PCR column purification (Qiagen). DNA fragmentation was controlled on a 2% agarose gel.

Sequencing Platform

instrument_model
Illumina Genome Analyzer IIx

hg38

Number of total reads
55876641
Reads aligned (%)
79.8
Duplicates removed (%)
24.2
Number of peaks
14705 (qval < 1E-05)

hg19

Number of total reads
55876641
Reads aligned (%)
78.7
Duplicates removed (%)
24.7
Number of peaks
15072 (qval < 1E-05)

Base call quality data from DBCLS SRA