GSM8970985: Male Wild-type Mouse 1; Mus musculus; RNA-Seq
Total RNA was isolated from homogenized pancreatic tissue by the guanidinium thiocyanate/phenol method according to Sparmann et al. (Sparmann G, Jäschke A, Loehr M, Liebe S, Emmrich J. Tissue homogenization as a key step in extracting RNA from human and rat pancreatic tissue. Biotechniques 1997; 22(3):408-10, 412.). Total RNA was quantified by spectrophotometry (NanoDrop 1000, Thermo Fisher Scientific, Waltham, MA, United States), and their integrity was checked using the Agilent Bioanalyzer 2100 with an RNA Nano chip kit (Agilent Technologies, Waldbronn, Germany). Afterwards, the samples were transferred to Novogene (Cambridge, UK) for further processing. Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers, followed by the second strand cDNA synthesis using either dUTP for directional library or dTTP for non-directional library. For the non-directional library, it was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. For the directional library, it was ready after end repair, A-tailing, adapter ligation, size selection, USER enzyme digestion, amplification, and purification.