title: Sample 163456
organism: Mus musculus
description: Protocols: Colonic tissue was cut into small pieces and incubated with complete media including antibiotics and antimycotics at 37°C for 15 min. Colonic tissue was washed twice and then incubated with an enzyme cocktail (trypsin (2.5 mg/ml; Difco), collagenaseIV (400 U/ml), DNase1 (350 U/ml) at 37°C for 30 min. The supernatant (containing cells in suspension) was removed and filled with complete media, and the remaining tissue was incubated with enzyme cocktail at 37°C for another 20-30 min. If the tissue was not completely disrupted by then, it was further disintegrated by passing through a 23G needle. Next, all cell suspensions were put together and filtered through a 100m cell strainer. Cells were pelleted by 1500 rpm for 5 min. After additional washes with complete media, cells were cultured in tissue culture flasks and media was renewed after 1 day and subsequently when necessary. Adhering fibroblasts grew to confluence within some days. Intestinal fibroblasts from the colons of C57BL/6 wild-type mice were grown in the presence or absence of IL-36ɣ (30 ng/ml) for 4 hours. The experimental setup included 6 samples from 3 colons in a pairwise design (3 stimulated vs 3 unstimulated). RNA was purified from colonic fibroblasts with PeqGold MicroSpin Total RNA Kit according to the manufacturers` protocol. Additional DNA removal was performed with DNA Removing Columns. The sequencing library was NuGEN Ovation RNA-Seq Library Prep-Kit.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2015-12-31T17:01:16Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-09T03:55:23Z
attributes.tag: External Id
attributes.value: SAMEA3501701
attributes.tag: INSDC center name
attributes.value: Universitaetsklinikum Erlangen Department of Medicine 1
attributes.tag: INSDC first public
attributes.value: 2015-12-31T17:01:16Z
attributes.tag: INSDC last update
attributes.value: 2018-03-09T03:55:23Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-3814:Sample 163456
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: colonic fibroblast
attributes.tag: common name
attributes.value: house mouse
attributes.tag: individual
attributes.value: 1
attributes.tag: sample name
attributes.value: E-MTAB-3814:Sample 163456
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: strain
attributes.value: C57BL/6