ChIP-Atlas: SRX277081

Antigen

Cell type Class: Blood
Cell type: Lymphocytes
MeSH Description: White blood cells formed in the body's lymphoid tissue. The nucleus is round or ovoid with coarse, irregularly clumped chromatin while the cytoplasm is typically pale blue with azurophilic (if any) granules. Most lymphocytes can be classified as either T or B (with subpopulations of each), or NATURAL KILLER CELLS.

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: Lymphocytes were isolated from 10 ml Heparin blood using a swelling buffer (155 mM NH4CL2, 10 mM KHCO3, 0,01 mM EDTA) to disrupt erythrocytes. Cells were cross-linked in 1% formaldehyde for 10 min at room temperature. Chromatin was sonicated using a Bioruptor sonicator (Diagenode, Liège, Belgium) for 28 minutes at high power, 30 s ON, 30 s OFF, and ChIP was performed using a CTCF antibody (Millipore, Billerica, MA, USA; 07-729). 7 ng of DNA as measured by Qubit (Life Technologies) was prepared for sequencing. A barcoded NEXTflex adaptor was ligated to the DNA fragments, and fragments with adaptors were amplified by a pre-size selection PCR of 4 cycles, followed by size selection (∼300 bp) and further amplification by 14 PCR cycles. Three barcoded samples were sequenced in one lane on a HiSeq 2000 (Illumina, San Diego, CA, USA). The reads were demultiplexed using CASAVA 1.8.2 (Illumina).