Sample information curated by ChIP-Atlas

Antigen

Antigen Class
Unclassified
Antigen
Unclassified

Cell type

Cell type Class
Others
Cell type
SCC-25
Primary Tissue
Tongue
Tissue Diagnosis
Carcinoma Squamous Cell

Attributes by original data submitter

Sample

source_name
cultured cells
biomaterial_provider
ATCC
cell line
SCC25
cell type
head and neck sequamous cell carcinoma cell line
experiment type
ChIP-seq
ip-target
mock_rabbit IgG

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
SCC25 and HGEP cells were fixed with 1% formaldehyde for 10 min. For histone ChIP-seq we used the Chromatin Shearing Optimization Kit-Low SDS (Diagenode) with 10 million cells per cell line and 40 cycles sonication (30 sec ON, 30 sec OFF, High) in the Bioruptor Plus (Diagenode). After RNase and proteinase K incubation, chromatin shearing was confirmed as a DNA smear between 100-300 bp on a 1.5% agarose gel. Histone ChIP DNA was prepared with the Auto Histone ChIP-seq kit protein G Kit (Diagenode), with 3 μg H3K4me1, 2 μg H3K4me3, 2.4 μg H3K27ac, 2 μg H3K27me3 and 2.4 μg H3K36me3 premium polyclonal antibody (Diagenode) per 1 million cells with the SX-8G IP-Star® Compact System (Diagenode). 1% input sample for each cell line was used as a control for background experimental noise. ChIP and input DNA were quantified with the Quant-iT PicoGreen ds DNA Assay Kit (Invitrogen). ChIP-seq libraries were made with the ThruPLEXTM-FD Prep Kit (Rubicon Genomics), purified and size-selected with a 2% agarose Pippin cassette (Sage Science). Libraries were quantified with the Quant-iT PicoGreen ds DNA Assay Kit (Invitrogen) and the KAPA Library Quantification Kit (Kapa Biosystems). Quality control was performed with the Agilent Technologies 2100 Bioanalyzer. Libraries were normalized based on qPCR and pooled with the TruSeq SR Cluster Kit v3-cBot-HS (Illumina). Pooled samples were multiplexed and sequenced with the HiSeq 2500 v3 (Illumina).

Sequencing Platform

instrument_model
Illumina HiSeq 2500

hg38

Number of total reads
19792981
Reads aligned (%)
95.0
Duplicates removed (%)
40.3
Number of peaks
669 (qval < 1E-05)

hg19

Number of total reads
19792981
Reads aligned (%)
94.1
Duplicates removed (%)
41.7
Number of peaks
766 (qval < 1E-05)

Base call quality data from DBCLS SRA