characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2014-12-23T17:01:35Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T22:39:58Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA3178837
characteristics.characteristic: External Id
characteristics.text: Imperial Centre for Translational and Experimental Medicine
characteristics.characteristic: INSDC center name
characteristics.text: 2014-12-23T17:01:35Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T22:39:58Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS631757
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-3199:Human Islet HI32
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: islet cell
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UBERON_0000016
characteristics.characteristic: cell type
characteristics.text: human
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: common name
characteristics.text: Protocols: Pancreatic Islets were isolated following the method of Bucher et al. (2005), Transplantation, 79: 91-97. Isolated islets were were cultured in CRML 1066 medium containing 10% FCS serum for shipping and, if not processed immediately, were recultured for 3 additional days. Cultured islets were rinsed with PBS buffer 3 times and cross-linked for 10 min in 1% formaldehyde at room temperature with constant shaking. After adding glycine (final concentration 125 mM), islets were rinsed with PBS-containing protease inhibitor cocktail (Roche) at 4 degreesC, snap frozen and stored at -80 degreesC. Islet purity was assessed by dithizone staining34 immediately before fixation. Protocol as described in Giresi et al. (2007), Genome Research, 17:877-855, with the following modifications: frozen pellets with ~3,000 cross-linked islets were thawed on ice in 1 ml lysis buffer (2% Triton X-100, 1% SDS buffer, 100 mM NaCl, 10 mM Tris-Cl at pH 8.0 and 1 mM EDTA buffer) and disrupted with five 1-min cycles using 0.5-mm glass beads (BioSpec). Samples were sonicated for 10-20 rounds of 30 pulses each (1 s on and 0.5 s off) using a Branson Sonifier 450D at 15% amplitude. After 10 rounds, the efficiency of sonication was assessed, and further rounds were performed when needed to ensure that the majority of chromatin fragments were in the 200-1,000-bp range. Debris was cleared by centrifugation at 15,000g for 5 min at 4 degreesC. Nucleosome-depleted DNA was extracted with phenol-chloroform followed by ethanol precipitation and RNase A (100 ?g/ml) treatment. Libraries were generated from gel-purified ~200-bp DNA fragments. Adapters were amplified using ligation-mediated PCR as detailed in Ren et al. (2000), Science, 290: 2306-2309.
characteristics.characteristic: description
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: Human Islet HI32
characteristics.characteristic: title