GSM4729333: STD-fed WT1; Mus musculus; RNA-Seq
G solution was prepared by adding 0.25% (wt./vol.) BSA into HBSS (Invitrogen, USA). Collagenase P (Roche, Switzerland) was dissolved in G solution to obtain 0.8 mg/ml collagenase solution. Ice cold collagenase solution (around 3 ml) was injected into the pancreas of anesthetized mouse through the joint site of the hepatic duct and the cystic duct of the common bile duct. Pancreas was removed and infused in another 3 ml 0.8 mg/ml ice cold collagenase solution in centrifuge tube. Pancreas was then transferred to a 37 ℃ water bath and gentle shaken (200 rpm) for 8 min. Pancreas was shaken vigorously for 10 s at the end of the digestion. Digested tissues were washed with 10 ml G solution three times and filtered with 500 µm mesh. Tissues were resuspended with G solution and gradient centrifuged with Histopaque 1119, 1083 and 1077 (Sigma-Aldrich, USA). Islet layer was poured into a 70 µm cell strainer. Finally, the strainer was turned upside down and rinsed with RPMI 1640 medium on a petri dish to capture the islets. RNA libraries were prepared for sequencing using standard Illumina protocols