7d Dox 3TF-1
Mus musculus
Protocols: Mice were treated with doxycycline (Sigma) beginning at 6 to 8 weeks of age. The drug was dissolved in a 5% sucrose solution and provided ad libitum in lieu of the normal water supply. Pancreata were removed, perfused with 2 mg/mL collagenase P (Roche) in HBSS, minced and incubated in 37ďžĄC for 5 minutes. The cells were further dispersed by manual pipetting, washed with FACS staining buffer (R&D Systems), filtered through a 100 _m mesh cell strainer then centrifuged. Cell pellets were resuspended in AccuMax (Sigma) and DNase1 (3U, Ambion) and incubated at 37ďžĄC for 4 minutes. Afterwards, cells were washed with FACS staining buffer, centrifuged and resuspended in FACS buffer containing DNAse1 (1 U/ml, Ambion) and 1mM EDTA. DAPI was used to stain for cell viability at a dilution of 1:1000. 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 using the Ovation RNA-seq system (Nugen, 7100-08). The library was constructed using the Illumina TruSeq DNA sample prep kits A&B, FC-121-1001 and FC-121-1002 with the standard Illumina methods (Bentley 2008). Median library insert length=155; adapter length=100; amount amplified=10 ng.
Characteristics [cell type
acinar cell
ENA-FIRST-PUBLIC
2016-07-06T17:01:44Z
ENA-LAST-UPDATE
2018-03-09T05:10:53Z
External Id
SAMEA3556806
INSDC center name
Vanderbilt University and University of Pennsylvania
INSDC first public
2016-07-06T17:01:44Z
INSDC last update
2018-03-09T05:10:53Z
INSDC status
public
Submitter Id
E-MTAB-3921:7d Dox 3TF-1
age
P60
broker name
ArrayExpress
common name
house mouse
sample name
E-MTAB-3921:7d Dox 3TF-1
scientific_name
Mus musculus
strain
Gt (ROSA)26tm5(3TF.mCherry)Mgn