Sample information curated by ChIP-Atlas

Antigen

Antigen Class
TFs and others
Antigen
DOT1L

Cell type

Cell type Class
Blood
Cell type
HCB1
NA
NA

Attributes by original data submitter

Sample

source_name
HCB1_ChIP_DOT1L_dVHL_500nM_3h_REP1_1909x2_TGATACGT_S22
cell line
HCB1
antibody
DOT1L
antibody manufacturer
CST
antibody catalog number
77087
agent
500nM dVHL
timepoint
3 hours

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
The cells were collected and washed twice in ice-cold PBS followed by crosslinking in 1% formaldehide (methanol free) for 7-8 minutes. Excess of formaldehyde was quenched by addition of Tris pH 8.0 to 100nM final and Glycine 125 mM final. Chromatin was sheared using Covaris E220 for 30 min with power 140; duty 5; bursts 200/sec. We then used a 5ul aliquot of sherared chromatin to decrosslink in order to test completeness of shearing. Chromatin passed QC if fragments between 100-700bp were >90%. For immunoprecipitation we used chromatin from 20x10^6 cells, 5ug of specific antibody and Protein-A/G magnetic beads (Dynal). Following roatation at 4 degrees overnight, the immune complexes were wased in Mixed Micelle Buffer, buffer 500, LiCl/detergent solution and TE buffers. DNA was eluted from the beads using 100mM NaHCO3 100mM NaCl 1% SDS and purified using AmpureXP beads. 1-10 ng of ChIP'ed DNA was used for the library preparation using the SMARTer ThruPLEX DNA-Seq Kit (Takara) Sequencing was done using the Illumina Next Gen Sequencing NextSeq platform (Illumina, San Diego, CA) with 37bp, paired-end reads.

Sequencing Platform

instrument_model
NextSeq 500

hg38

Number of total reads
37424226
Reads aligned (%)
98.9
Duplicates removed (%)
11.9
Number of peaks
1331 (qval < 1E-05)

hg19

Number of total reads
37424226
Reads aligned (%)
97.9
Duplicates removed (%)
12.1
Number of peaks
1164 (qval < 1E-05)

Base call quality data from DBCLS SRA