organism: Mus musculus
description: Protocols: Mouse embryonic stem cells (mES) were detached from gelatinesed flasks by addition of trypsin. Cells were disassembled from 3-dimensional gastruloids (initiated from mES cells and biased towards hemato-endothelial cell production) at endopoint (216 h) using the dissociation agent TrypLE Express. Cells suspensions were quenched with medium and centrifuged at 400 x g for 5 minutes to pellet for subsequent extraction. Total RNA was extracted using Trizol (Invitrogen) and purified using Direct-zol RNA Purification Kits (Zymo Research). Library construction was performed by Novogene UK (Cambridge, UK). A minimum of 400 ng (20 ng/μl) of high-quality total RNA (extracted using Trizol, Invitrogen, and purified using Direct-zol RNA Purification Kits, Zymo Research) was supplied for sequencing. Novogene UK constructed cDNA libraries by purification of messenger RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, PCR amplification, and purification.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2022-10-12
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2022-10-20
attributes.tag: External Id
attributes.value: SAMEA111349844
attributes.tag: INSDC center alias
attributes.value: Brunel University London
attributes.tag: INSDC center name
attributes.value: Brunel University London
attributes.tag: INSDC first public
attributes.value: 2022-10-12T09:19:55Z
attributes.tag: INSDC last update
attributes.value: 2022-10-20T16:34:51Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-12173:mES_EV_rep2
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell_line
attributes.value: embryonic cell line
attributes.tag: cell_type
attributes.value: embryonic stem cell
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: embryo stage
attributes.tag: genotype
attributes.value: empty vector
attributes.tag: growth condition
attributes.value: normal cell culture
attributes.tag: organism part
attributes.value: embryoid body
attributes.tag: sample name
attributes.value: E-MTAB-12173:mES_EV_rep2