GSM7659973: 15861 TP53 5 A31; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
TFs and others
Antigen
TP53
Cell type
Cell type Class
Blood
Cell type
Acute myeloid leukemia
NA
NA
Attributes by original data submitter
Sample
source_name
primary patient
cell type
AML
cell line
NA
antibody
Santa Cruz: DO-1, sc-126X
icu admission
NA
chip target
ChIP-seq TP53
geo_loc_name
missing
collection_date
missing
Sequenced DNA Library
library_name
GSM7659973
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Primary AML cells and cell lines were crosslinked with 1% formaldehyde for 10 min at room temperature at a concentration of 30 × 106 cells/mL. The fixed cells were sonicated 10–15 cycles (30 sec on, 30 sec off) to a size of 200–600 bp. Sonicated chromatin was incubated overnight with either 5 µg TP53 antibody (Santa Cruz: DO-1, sc-126X) or 2 µg of antibody against H3K4me3, H3K27me3, or H3K27ac. The next day, antibody–protein complexes were bound to magnetic Protein G beads (Invitrogen, ThermoFisher, Netherlands) for ∼2 hours. Beads were washed with four different wash buffers, and chromatin was eluted from the beads. DNA proteins were de-crosslinked, and samples were purified using the Qiaquick MinElute PCR purification kit. Sequencing samples were prepared according to the manufacturer's protocol (Illumina, Eindhoven, Netherlands). End repair was performed using the precipitated DNA using Klenow and T4 PNK, and subsequently a 3′ protruding A base was generated using Taq polymerase followed by adapter ligation. The DNA was amplified by PCR and ∼300 bp (ChIP fragment + adapters). Samples were sequenced on an Illumina NextSeq 500 system with 2×43 bp paired-end sequencing (PE43).