title: Cells from pool of 46 E10.5 Ngn3EGFP/+ [het] embryos (13 litters)
organism: Mus musculus
description: Protocols: Manual dissection was employed to remove as much non-pancreatic tissue as possible. Tissues were dissociated with 0.025% trypsin to mostly single cells. Then the following steps were perfomed: (i) neutralize with 5% FBS, (ii) spin to remove solution, (iii) resuspend and sort cells into 2%FBS in PBS. Purity was visually inspected to verify EGFP fluorescence and viability and it was estimated by by cytospin. Total RNA was isolated using TRIzol LS (Invitrogen) and column purification (Zymo Research), according to manufacturer's instructions. RNA was DNase-treated (Ambion) and integrity was assessed on the Bioanalyzer (Agilent). Total RNA was amplified (0.5-10ng) using the Ovation RNA-Seq system (NuGEN). The transcriptional sequencing methods were performed essentially as previously described (Mortazavi 2008), which is a modification of the standard Illumina methods (Bentley 2008). Median fragment length=700, amount amplified = 5 ng.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2015-01-26T17:02:21Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-08T22:55:15Z
attributes.tag: External Id
attributes.value: SAMEA3220365
attributes.tag: INSDC center name
attributes.value: Vanderbilt University
attributes.tag: INSDC first public
attributes.value: 2015-01-26T17:02:21Z
attributes.tag: INSDC last update
attributes.value: 2018-03-08T22:55:15Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-3272:Cells from pool of 46 E10.5 Ngn3EGFP/+ [het] embryos (13 litters)
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: Ngn3-expressing cell
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: embryonic day 10.5 (E10.5)
attributes.tag: sample name
attributes.value: E-MTAB-3272:Cells from pool of 46 E10.5 Ngn3EGFP/+ [het] embryos (13 litters)
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: strain
attributes.value: B6.129S-Neurog3tm1(EGFP)Khk/Mmcd