title: Sample 11
organism: Mus musculus
description: clone13-4-2ibulk 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 1 uM 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 mRNA libraries were prepared using TruSeq RNA Sample Preparation (Illumina) following the manufacturer's instructions.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2015-12-31T17:01:13Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-08T19:45:02Z
attributes.tag: External Id
attributes.value: SAMEA2697847
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: 2018-03-08T19:45:02Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-2830:Sample 11
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: house mouse
attributes.tag: growth condition
attributes.value: 2i
attributes.tag: sample name
attributes.value: E-MTAB-2830:Sample 11
attributes.tag: scientific_name
attributes.value: Mus musculus