GSM5456904: SUDLH5 EV RAD21 ChipSeq; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
TFs and others
Antigen
RAD21
Cell type
Cell type Class
Blood
Cell type
SU-DHL-5
Primary Tissue
Blood
Tissue Diagnosis
Lymphoma B-cell
Attributes by original data submitter
Sample
source_name
SU-DHL-5
treatment
empty vector
sirna
-
antibody
RAD21 (Bethyl Laboratories, A300-080A)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cells were crosslinked with 1% formaldehyde for 5 min. 6% murine T-lymphoma cells were added during cell lysis as exogenous spkie-in for normalization. After cell lysis, nuclei were re-suspended in RIPA buffer (10 mM Tris/HCl pH 7.5, 150 mM NaCl, 1% NP40, 1% deoxycholic acid (DOC), 0.1% SDS, 1 mM EDTA) and DNA was fragmented to a size <300 bp using a Branson sonifier or using the Covaris Focused Ultrasonicator M220 for 100 min per ml lysate. Chromatin was immunoprecipitated with 15 µg of corresponding antibody for 6 to 8 hours. Chromatin was eluted with 1% SDS and crosslinking was reverted overnight. For purification, chloroform/phenol extraction was used. NEBNext® ChIP-Seq Library Prep Master Mix Set for Illumina® or NEBNext® Ultra II DNA Library Prep with Sample Purification Beads (New England Biolabs). Purified DNA was end-repaired, A-tailed, ligated to Illumina adaptors, size-selected (200 bp) and purified with Qiagen gel extraction kit or Sample Purification Beads. DNA fragments were amplified by 17 cycles of PCR and library size was tested with the Fragment Analyzer (Advanced Analytical). The amount of library DNA was quantified using a picogreen assay.