Essential Role of Thioredoxin and Glutathione in Pancreas
Essential Role of Thioredoxin and Glutathione in Pancreas
Pancreatic tissue was collected from 8-12 week old mice Acute pancreatitis was induced as published (Algül et al., JCI 2007), Buthionine sulphoximine (BSO) was given at 20mM in the drinking water  for 48 hours before AP induction Maxwell 16 Tissue LEV Total RNA Purification Kit was used for RNA extraction according to the manufacturer's instructions (Promega) Library preparation for bulk 3'-sequencing of poly(A)-RNA was done as described previously (Parekh et al., 2016) barcoded cDNA of each sample was generated with a Maxima RT polymerase (Thermo Fisher) using oligo-dT primer containing barcodes, unique molecular identifiers (UMIs) and an adapter. 5' ends of the cDNAs were extended by a template switch oligo (TSO) and after pooling of all samples full-length cDNA was amplified with primers binding to the TSO-site and the adapter. cDNA was tagmented with the Nextera XT kit (Illumina) and 3'-end-fragments finally amplified using primers with Illumina P5 and P7 overhangs.