Homo sapiens
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.
ENA-FIRST-PUBLIC
2018-08-28T17:03:20Z
ENA-LAST-UPDATE
2018-07-20T15:42:30Z
External Id
SAMEA4799853
INSDC center name
IBMC/i3S
INSDC first public
2018-08-28T17:03:20Z
INSDC last update
2018-07-20T15:42:30Z
INSDC status
public
Submitter Id
E-MTAB-6661:Sample 2
broker name
ArrayExpress
cell line
HeLa
cell type
epithelial cell
common name
human
disease
cervical adenocarcinoma
organism part
uterine cervix
sample name
E-MTAB-6661:Sample 2
scientific_name
Homo sapiens