title: Sample 12
organism: Mus musculus
description: clone13-4-high-IDE1-d3 Protocols: Mouse ESCs were maintained without feeders in 'LIF+serum' medium (DMEM (high glucose, no glutamine, with sodium bicarbonate, Invitrogen) supplemented with 15% ES-qualified EmbryoMax Fetal Calf Serum (Millipore), 10 ng/ml murine LIF (EMBL Protein Expression and Purification Core Facility), 1x Non-Essential Amino Acids, 2 mM L-glutamine, 1 mM sodium pyruvate, 100 U/ml penicillin and 100 ug/ml strep- tomycin, 0.1 mM 2-mercaptoethanol (all Invitrogen) on culture dishes (Nunc) coated with 0.1% gelatin (Sigma) solution and cultured at 37C with 5% CO2. Medium was changed daily and cells were passaged every other day with 0.05% Trypsin-EDTA (Invitrogen) at a passaging ratio of 1/6- 1/12. N2B27 medium was prepared from a 1:1 mixture of DMEM/F12 (without HEPES, with L-glutamine) and neurobasal medium with 0.5x B-27 (without vitamin A) and 0.5x N-2 supplements, 100 U/ml penicillin and 100 ug/ml streptomycin, 0.25 mM L-glutamine, 0.1 mM 2-mercaptoethanol (all Invitrogen), 10 ug/ml BSA fraction V and 10 ug/ml human recombinant insulin (both Sigma). '2i' medium was N2B27 medium supplemented with 3 uM CHIR99021 and 1uM PD0325901 (both Tocris Bioscience). mESCs were differentiated to endoderm precursors in Advanced RPMI, 100 U/ml penicillin and 100 ug/ml streptomycin (both Invitrogen), 0.2% FCS (EmbryoMax, Millipore), 1 uM IDE-1 (Tocris) following Borowiak et al. (Cell Stem Cell 2009). Barcoded miRNA libraries were prepared using NEBNext Multiplex Small RNA Library Prep Set for Illumina (New England Biolabs) following the manufacturers instructions
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2015-12-31T17:01:13Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-08T19:34:13Z
attributes.tag: External Id
attributes.value: SAMEA2698008
attributes.tag: INSDC center name
attributes.value: European Molecular Biology Laboratory
attributes.tag: INSDC first public
attributes.value: 2015-12-31T17:01:13Z
attributes.tag: INSDC last update
attributes.value: 2018-03-08T19:34:13Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-2831:Sample 12
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: house mouse
attributes.tag: growth condition
attributes.value: IDE1 endodermal differentiation
attributes.tag: sample name
attributes.value: E-MTAB-2831:Sample 12
attributes.tag: scientific_name
attributes.value: Mus musculus