title: Donor 04_occipital
organism: Homo sapiens
description: Protocols: 10 pilosebaceous units (PUs) at the anagen phase were isolated from each involved (balding; vertex) and non-involved (control; occipital) scalp region using the follicular unit extraction (FUE) technique. Total RNA was extracted from hair tissue stabilized in RLT buffer according to the “Purification of total RNA from animal and human tissue” protocol of the RNeasy Micro Kit (QIAGEN, Hilden, Germany). In brief, the hair tissue was stored in RLT buffer containing β-mercaptoethanol and shipped on dry ice. The samples were homogenized in RLT buffer using Precellys CK14 ceramic beads (2 cycles of 20 seconds at 6500 rpm; 15 seconds break). Next one volume of 70 % ethanol was added and the samples were applied to RNeasy MinElute spin columns followed by an on-column DNase digestion and several wash steps. Finally total RNA was eluted in 14 μl of nuclease free water. Purity and integrity of the RNA was assessed on the Agilent 2100 Bioanalyzer with the RNA 6000 Nano LabChip reagent set (Agilent, Palo Alto, CA, USA). In brief, 75 ng of total RNA were reverse transcribed into first strand cDNA using a mixture of random and poly-dT primers, omitting the integrated DNase treatment step. Second strand synthesis, using a nucleotide analog enabling strand retention, generated double stranded cDNA (ds cDNA). The ds cDNA was fragmented to an average size of 200-500 bases using a Covaris S2 Sonication System (Intensity 5; Duty Cycle 10%; Cycles per Burst 200; Treatment Time 120 s; micro Tube AFA Fiber Snap-Cap). Next, an end repair was performed to generate blunt-ended ds cDNA, followed by the ligation of the indexing adapters, a strand selection via nucleotide analog-targeted degradation and a reduction of the rRNA content by an InDA-C-mediated adaptor cleavage. Finally cDNA libraries were created by a 18 cycles PCR enrichment. The libraries were quantified using the KAPA SYBR FAST ABI Prism Library Quantification Kit.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2020-09-20T04:05:04Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2020-09-10T15:53:23Z
attributes.tag: External Id
attributes.value: SAMEA7301447
attributes.tag: INSDC center name
attributes.value: L'Oreal Research & Innovation Advanced Research
attributes.tag: INSDC first public
attributes.value: 2020-09-20T04:05:04Z
attributes.tag: INSDC last update
attributes.value: 2020-09-10T15:53:23Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-9557:Donor 04_occipital
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: disease
attributes.value: androgenetic alopecia
attributes.tag: individual
attributes.value: Donor 04
attributes.tag: organism part
attributes.value: hair follicle
attributes.tag: sample name
attributes.value: E-MTAB-9557:Donor 04_occipital
attributes.tag: sampling site
attributes.value: occiput
attributes.tag: scientific_name
attributes.value: Homo sapiens
attributes.tag: sex
attributes.value: male