Sample information curated by ChIP-Atlas

Antigen

Antigen Class
TFs and others
Antigen
FOS

Cell type

Cell type Class
Pancreas
Cell type
CFPAC-1
Primary Tissue
Pancreas
Tissue Diagnosis
Adenocarcinoma Ductal

Attributes by original data submitter

Sample

source_name
CFPAC1.ChIP.FOS
cell line
original CFPAC1 cell line
cell type
Pancreatic Ductal Adenocarcinoma (PDAC) cell line
antibody
Anti-FOS (Antibody Sigma HPA018531 - Lot E106326)

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
ChIP lysates were generated from 50x10^6 cells. Cells were either fixed in 1% formaldehyde for 10min (for FOXA2, HOXB8, JUNB and FOXA3 ChiP-seq) or with a double cross-linking protocol consisting in 45 minutes of 2 mM DSG followed by 10 minutes in 1% formaldehyde for FOS ChIP-seq. Lysate was immunoprecipitated with 10ug of antibody. Antibodies were pre-bound overnight to 100ul 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 was prepared for HiSeq 2000 Illumina sequencing using a standard protocol consisting in blunting, addition of dA overhangs, ligation of Illumina adapters, PCR with index primers and purification. A mixture of T4 DNA polymerase, DNA polymerase I and T4 kinase was used according to manufacturer's instruction. Library preparation is carried out on SPRIworks Fragment Library System.

Sequencing Platform

instrument_model
Illumina Genome Analyzer II

hg19

Number of total reads
35729181
Reads aligned (%)
65.2
Duplicates removed (%)
32.2
Number of peaks
23570 (qval < 1E-05)

hg38

Number of total reads
35729181
Reads aligned (%)
66.2
Duplicates removed (%)
31.4
Number of peaks
23711 (qval < 1E-05)

Base call quality data from DBCLS SRA