title: DUY276A3
organism: Homo sapiens
description: Protocols: Whole skin biopsies were kept on ice immediately after resection from facial skin for no longer than 2hrs before processing. The skin samples were either directly processed for dissociation into cell suspension or kept in MACS Tissue Storage Solution (Miltenyi Biotec, Surrey, U.K, cat. no. 130-100-008) overnight. The enzymatic and mechanical dissociation of whole skin biopsies was performed using the Whole Skin Dissociation kit for human material (Miltenyi Biotec, Surrey, U.K, cat. no. 130-101-540). Larger specimens were reduced in size by pre-cutting into several small pieces prior to enzymatic digestion and the quantity of enzymes used was adjusted according to the size of the sample. All biopsies were incubated with the enzyme solution for 3 hours in a shaking water bath at 37°C. Cell suspensions were then filtered through 70-µm cell strainer (Falcon) and centrifuged at 1500rpm for 10 minutes at 4°C. The collected cells were then frozen in 10%DMSO in Fetal Bovine Serum (FBS) and stored in liquid nitrogen. For library preparation, cells were thawed and resuspended in PBS with 2% FBS and DAPI (0.05mg/ml). Dead cells were removed through flow cytometry of the final cell suspension. Immediately after flow cytometry, ~20,000 to 100,000 live single cells were loaded onto a Chromium chip and libraries were prepared through droplet encapsulation on the Chromium controller (10X Genomics; Pleasanton, CA, USA) using the Single Cell 3' reagent kits following the manufacturer's protocol. Library quantification was performed using the Qubit dsDNA HS Assay Kit (Life Technologies, Waltham, MA, USA), and cDNA integrity was assessed using D1000 ScreenTapes (Agilent Technologies).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2023-12-22T00:41:15Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2023-12-22T00:41:15Z
attributes.tag: External Id
attributes.value: SAMEA115054637
attributes.tag: INSDC center name
attributes.value: Centre for Gene Therapy and Regenerative Medicine, King's College London, London, UK -Wellcome Sanger Institute, Cambridge, UK
attributes.tag: INSDC first public
attributes.value: 2023-12-22T00:41:15Z
attributes.tag: INSDC last update
attributes.value: 2023-12-22T00:41:15Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-13085_3:DUY276A3
attributes.tag: age
attributes.value: 54
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell_type
attributes.value: skin 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: normal
attributes.tag: geographic location (country and/or sea)
attributes.value: not collected
attributes.tag: individual
attributes.value: human whole skin biopsy
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: skin
attributes.tag: sample id
attributes.value: PSU_microdissected_data
attributes.tag: sample name
attributes.value: E-MTAB-13085_3:DUY276A3
attributes.tag: sampling site
attributes.value: scalp
attributes.tag: scientific_name
attributes.value: Homo sapiens
attributes.tag: sex
attributes.value: female