title: Illumina HiSeq 2000 sequencing; RNA-Seq of isolated pancreatic stellate cells from mice with autoimmune pancreatitis and chronic pancreatitis
design: RNA-Seq of isolated pancreatic stellate cells from mice with autoimmune pancreatitis and chronic pancreatitis
library_name: 129_s
library_construction_protocol: Pancreas tissue was collected from the animals sacrificed at the end of the experiment. Isolation of the pancreatic stellate cells (PSCs) was performed by gradient centrifugation using Nycodenz solution. The cells layer was collected and purified by negative selection using CD45 microbeads and MACS LD column.  To induce pancreatitis, animals were injected with three episodes of caerulein (50Î¼g/kg six time each episode) per week over two weeks. To activate the cre recombinase, animals received daily tamoxifen treatment for 5 days during the first week. All animals were sacrificed 3 weeks after the first injection episode. The total RNA from cells was extracted with the RNeasy Mini kit (Qiagen) according to the manufacturer's protocol. Library preparation: Random primed cDNA library- Purification of poly-A containing mRNA molecules- mRNA fragmentation- Random primed cDNA synthesis- Adapter ligation and adapter specific PCRamplification
attributes.tag: Experimental Factor: phenotype
attributes.value: wild type phenotype
attributes.tag: Experimental Factor: disease
attributes.value: chronic pancreatitis