title: Replicate 2 DG75 parental ChIP
organism: Homo sapiens
description: Protocols: For immunoprecipitation two samples with 1×108 cells each were adjusted to a concentration of 5×105 cells/ml in fresh medium and were left non-induced or were induced with a final concentration of 100 ng/ml doxycycline (Sigma-Aldrich) for 15 h. 1 x 10^8 cells were adjusted to a concentration of 5 x 10^5 cells/ ml prior to chromatin preparation. Nuclei were extracted with hypotonic buffer (10 mM KCl, 340 mM Sucrose, 1.5 mM MgCl2, 10 mM Hepes pH 7.9) containing 10 % proteinase inhibitor cocktail (PIC, Roche) and lysed in RIPA buffer + 1x PIC. The chromatin was shared in a BioRuptor on wet ice (4 cycles, 5 min each, 30 on/off, high). The chromatin was immunoprecipitated with the BZ1 antibody (Young et al., 1991), 10 % input was used as a control. The precipitates were washed with different salt concentrations and the proteins were digested with proteinase K. The library construction was performed with the NEBNext Ultra II DNA Library Prep Kit for Illumina (NWB # E7645S) and NEBNext Multiplex Oligos for Illumina (Index Primer Set 2, NEB #E7500S) starting from 8 ng DNA.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2019-04-05T04:02:49Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-03-26T23:44:20Z
attributes.tag: External Id
attributes.value: SAMEA5537264
attributes.tag: INSDC center name
attributes.value: Research Unit Gene Vectors Helmholtz Zentrum Munchen German Research Center for Environmental Health and German Center for Infection Research (DZIF), Partner site Munich, Germany
attributes.tag: INSDC first public
attributes.value: 2019-04-05T04:02:49Z
attributes.tag: INSDC last update
attributes.value: 2019-03-26T23:44:20Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-7821:Replicate 2 DG75 parental ChIP
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: DG-75
attributes.tag: cell type
attributes.value: B cell
attributes.tag: common name
attributes.value: human
attributes.tag: disease
attributes.value: Burkitts lymphoma
attributes.tag: sample name
attributes.value: E-MTAB-7821:Replicate 2 DG75 parental ChIP
attributes.tag: scientific_name
attributes.value: Homo sapiens