title: Sample 2
organism: Homo sapiens
description: Protocols: Mitotic extracts were derived from isolated mitotic cells treated cells with DMSO (control) or ActD (ActD). Total RNA was isolated from mitotic HeLa cells (shake-off) using trizol (Life technologies). Quantity and total RNA integrity was checked following isolation using an Agilent Technologies 2100 Bioanalyzer. Sample with RNA Integrity Number (RIN) value greater than 8 were selected. The RNA samples were prepared as described before in (Derks et al., 2015) with the following modifications. Five micrograms of total RNA were depleted according to the Illumina Ribo-zero magnetic protocol (www.illumina.com). The Ribo-zero treated RNA was purified using Agencourt RNAClean XP Beads. RNA was fragmented on a Covaris S220. One microliter of the sheared rRNA depleted RNA was loaded on an Agilent Technologies 21000 Bioanalyzer RNA Pico chip to determine successful ribosomal RNA depletion. The sheared RNA sample was concentrated in a Thermo scientific SPD100 speedvac to 5 µl and a sequencing library was prepared according to the Illumina TruSeq Small RNA protocol (www.illumina.com). In short, adapters are subsequently ligated to the 3'end and the 5'end of the RNA. The RNA is reverse transcribed, amplified by PCR and run on SDS-PAGE gel. RNA fragments between 140 - 500 bp are cut out of the gel and purified. One microliter was loaded on an Agilent Technologies 2100 Bioanalyzer using a DNA 1000 assay to determine the library concentration and quality.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2018-08-28T17:03:20Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-07-20T15:42:30Z
attributes.tag: External Id
attributes.value: SAMEA4799853
attributes.tag: INSDC center name
attributes.value: IBMC/i3S
attributes.tag: INSDC first public
attributes.value: 2018-08-28T17:03:20Z
attributes.tag: INSDC last update
attributes.value: 2018-07-20T15:42:30Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-6661:Sample 2
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: HeLa
attributes.tag: cell type
attributes.value: epithelial cell
attributes.tag: common name
attributes.value: human
attributes.tag: disease
attributes.value: cervical adenocarcinoma
attributes.tag: organism part
attributes.value: uterine cervix
attributes.tag: sample name
attributes.value: E-MTAB-6661:Sample 2
attributes.tag: scientific_name
attributes.value: Homo sapiens