title: PNUTS_IAA_rep2
organism: Homo sapiens
description: Protocols: Cells were treated with 2 μg/ml doxycycline and 0.06 μg/ml nocodazole, 24 and 18 h respectively, before the addition of 1 mM IAA to induce degradation of mAID-tagged proteins. After 4 h treatment with IAA, mitotic cells were manually detached by shake-off, and cells were transferred to new poly-L-lysin coated dishes. Once attached, cells were washed 3x with PBS and released from mitosis into G1 for 18 h in a fresh medium containing 2 mM thymidine.Cells were treated 30min with DNAseI, washed with PBS, detached with TrypLE, counted and pelleted by centrifugation. Cell pellet containing 100 000 cells was sent in dry-ice package to Active Motif S.A. Cells are lysed with a nonionic detergent to yield pure nuclei.  The resulting chromatin is then fragmented and simultaneously tagmented with sequencing adapters using the Tn5 transposase to generate the ATAC-Seq library. After purification, the library can be amplified by PCR using barcoded primers.
attributes.tag: ENA first public
attributes.value: 2024-01-01
attributes.tag: INSDC center name
attributes.value: College of Medicine, Health and Life Science, Brunel University London, Center of Genomic Engineering and Maintenance (CenGem)
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-13826:PNUTS_IAA_rep2
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell_line
attributes.value: HCT116
attributes.tag: cell_type
attributes.value: epithelial cell
attributes.tag: collection date
attributes.value: not collected
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: disease
attributes.value: colorectal carcinoma
attributes.tag: genotype
attributes.value: AID-degron PNUTS
attributes.tag: geographic location (country and/or sea)
attributes.value: not collected
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: colon
attributes.tag: sample name
attributes.value: E-MTAB-13826:PNUTS_IAA_rep2
attributes.tag: scientific_name
attributes.value: Homo sapiens