RNA-seq for isolated islets
Total RNA was extracted using TRIzol Reagent (TransGen Biotech, ET111-01) according to the manufacturers instructions. Genomic DNA was removed by digestion with DNase I. Ribosomal RNA (rRNA) was depleted by hybridizing complementary DNA oligonucleotides to rRNA, followed by selective degradation of RNA:DNA hybrids using RNase H (Beyotime, D7089). The resulting purified messenger RNA (mRNA) was fragmented into short fragments and subsequently used for the preparation of sequencing libraries.