title: mESc_MB_X2
organism: Mus musculus
description: Protocols: To induce differentiation and the exit from pluripotency mECs originating from 2iLIF culture were plated on gelatine-coated culture-dishes at a density of about 40.000/square cm in plain N2B27 medium (or 2iLIF medium as control) and cultured for 48h. Expression of constructs was induced by adding 1ug/ml doxycycline at the time of seeding. 1e5 to 1e7 cells were pelleted via centrifugation and total RNA was extracted with the mirVana miRNA Isolation Kit (Thermofisher) according to the manufacturers recommendations. Barcoded stranded mRNA-seq libraries were prepared from high quality total RNA samples (about 250 ng/sample) using the NEBNext Poly(A) mRNA Magnetic Isolation Module and NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs (NEB), Ipswich, MA, USA) implemented on the liquid handling robot Beckman i7. Obtained libraries that passed the QC step were pooled in equimolar amounts; 2.2 pM solution of this pool was loaded on the Illumina sequencer NextSeq 500 and sequenced uni-directionally, generating ~500 million reads, with an average read length of about 80 bases.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2020-11-10T04:11:06Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2020-07-23T12:16:56Z
attributes.tag: External Id
attributes.value: SAMEA7086909
attributes.tag: INSDC center name
attributes.value: EMBL
attributes.tag: INSDC first public
attributes.value: 2020-11-10T04:11:06Z
attributes.tag: INSDC last update
attributes.value: 2020-07-23T12:16:56Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-9404:mESc_MB_X2
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: embryonic stem cell
attributes.tag: common name
attributes.value: house mouse
attributes.tag: genotype
attributes.value: Rex1-GFPd2 iMC-linker (MN2_F5)
attributes.tag: growth condition
attributes.value: N2B27 2iLIF
attributes.tag: sample name
attributes.value: E-MTAB-9404:mESc_MB_X2
attributes.tag: scientific_name
attributes.value: Mus musculus