organism: Homo sapiens
description: Protocols: Human dermal fibroblasts (HDFs) retrieved from skin samples of neonatal (No. DFM021711A, Zen Bio) and octogenarian (AG10884, Coriell Institute) Caucasian males were used. All donors were reported as “healthy”. For all experiments, HDFs were seeded at 1x104 cells per cm2 of growth area in minimal essential medium Eagle-Earle (MEM) supplemented with 15% fetal bovine serum (FBS), 2mM L-glutamine, and 1x antibiotic-antimycotic (all from Gibco, Thermo Fisher Scientific). Only early passage dividing fibroblasts (up to passage 3-5) with cumulative population doubling level PDL < 24 were used. Total RNA from asynchronous cell populations was extracted using RNeasy® Mini Kit (Qiagen). 10ng were reverse-transcribed using the AmpliSeq Whole Transcriptome primers with the included SuperScript VILO cDNA Synthesis kit (Thermo Fisher Scientific). cDNA was used for target amplification (12 cycles) with Ion AmpliSeq primers and technology. Barcoded adapters were added and ligated to individual reactions according to the instructions of Ion AmpliSeq™ Transcriptome Human Gene Expression Kit (Thermo Fisher Scientific). The pooled libraries were processed on Ion Chef™ System(Thermo Fisher Scientific).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2020-02-27T04:05:19Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2020-01-06T13:11:02Z
attributes.tag: External Id
attributes.value: SAMEA6451245
attributes.tag: INSDC center name
attributes.value: Ipatimup
attributes.tag: INSDC first public
attributes.value: 2020-02-27T04:05:19Z
attributes.tag: INSDC last update
attributes.value: 2020-01-06T13:11:02Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-8634:N_Kif2B_1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: DFM021711A
attributes.tag: cell type
attributes.value: fibroblast of dermis
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: neonatal
attributes.tag: disease
attributes.value: normal
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: individual
attributes.value: DFM021711A
attributes.tag: organism part
attributes.value: zone of skin
attributes.tag: sample name
attributes.value: E-MTAB-8634:N_Kif2B_1
attributes.tag: scientific_name
attributes.value: Homo sapiens