organism: Homo sapiens
description: Protocols: Cells were harvested via trypsinization, followed by a series of wash steps. 100,000 cells of each were spun down at 550xg for 5min, 4C. Supernatant was removed; 50uL of PBS was added to wash pellet. Spin conditions remained the same. The cell pellet was resuspended in 50ul cold lysis buffer (1 mM Tris-HCl, pH 7.4, 10mM NaCl, 3mM MgCl2, 0.1% IGEPAL CA-630), spun down immediately at 550xg for 10min, 4C. The nuclei resuspended in the transposition reaction mix (2x TD Buffer (Illumina Cat #FC-121-1030, Nextera), 2.5ul Tn5 Transposes (Illumina Cat #FC-121-1030, Nextera) and Nuclease Free H2O) on ice. The transposition reaction was incubated at 37C for 45min. The transposed DNA was then purified using a Qiagen MinElute Kit with 10.5ul elution buffer. The transposed library was then PCR amplified using Nextera primers for 12 cycles. The PCR reaction was subsequently cleaned up using AMPureXP beads (Agencourt).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2019-04-30T04:02:46Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-12-17T11:56:33Z
attributes.tag: External Id
attributes.value: SAMEA5181580
attributes.tag: INSDC center name
attributes.value: The Children's Hospital of Philadelphia
attributes.tag: INSDC first public
attributes.value: 2019-04-30T04:02:46Z
attributes.tag: INSDC last update
attributes.value: 2018-12-17T11:56:33Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-7540:Sample 3
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: EndoC-betaH1
attributes.tag: cell type
attributes.value: beta cell
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: late embryo
attributes.tag: disease
attributes.value: normal
attributes.tag: organism part
attributes.value: pancreas
attributes.tag: replicate
attributes.value: 3
attributes.tag: sample name
attributes.value: E-MTAB-7540:Sample 3
attributes.tag: scientific_name
attributes.value: Homo sapiens