Sample information curated by ChIP-Atlas

Antigen

Antigen Class
TFs and others
Antigen
Pdx1

Cell type

Cell type Class
Pancreas
Cell type
NB490
NA
NA

Attributes by original data submitter

Sample

source_name
NB490 cell line
cell line
NB490
chip antibody
PDX1 (Millipore 07-696)
sample type
PDX1 ChIP

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Chromatin was prepared from the NB490 cell line using the truChIP Chromatin Shearing Reagent Kit (Covaris;520154). Chromatin was sheared in 1 mL millitubes (Covaris;520135) using a Covaris S200 (5% duty cycle, intensity of 4, 200 cycles per burst, for 8 minutes) to obtain chromatin fragments 100-700 base-pairs in length. 50 ug of fragmented chromatin was incubated overnight with 2.5 uL of rabbit anti-PDX1 antibody (Millipore;07-696). Complexes were isolated using 50 uL of Dynabeads (Invitrogen;10004D) previously blocked with BSA (1 mg/mL). 2 ng of immuno-precipitated DNA was prepared as a standard Illumina library using the ThruPLEX DNA-seq Kit (Rubicon), according to manufacturer's protocol. Samples were PCR-amplified and pooled. Final libraries were checked for quality and quantity by TapeStation (Agilent) and qPCR using Kapa?s library quantification kit for Illumina Sequencing platforms (Kapa Biosystems;KK4835). They were clustered on the cBot (Illumina) and sequenced 3 samples per lane on a 50-cycle single-end on a HiSeq 2000 in High Output mode using version 3 reagents.

Sequencing Platform

instrument_model
Illumina HiSeq 2000

mm10

Number of total reads
54947461
Reads aligned (%)
97.8
Duplicates removed (%)
16.0
Number of peaks
670 (qval < 1E-05)

mm9

Number of total reads
54947461
Reads aligned (%)
97.6
Duplicates removed (%)
16.0
Number of peaks
713 (qval < 1E-05)

Base call quality data from DBCLS SRA