title: SLIC-CAGE of paired-end sequencing data in mouse islets
design: SLIC-CAGE of paired-end sequencing data in mouse islets
library_construction_protocol: Mouse islets were isolated using collagenase (Roche) digestion as previously described(22). Islets were allowed to recover overnight preceding SLIC-CAGE and miRNA RT-qPCR experiments in RPMI 1640, 10% fetal bovine serum (FBS), L-glutamine, and 11 mM glucose at 37°C with 95% O2/ 5% CO2.  RNA was extracted using Trizol (Thermo Fisher Scientific) following the manufacturer's instructions. MiRNA reverse transcription (RT) and qPCR was performed from 30ng RNA as previously described(23) with miRCURY LNA probes and miRCURY LNA RT and SYBR® Green PCR Kits.  Let-7d-3p and miR-573-3p (data not shown) were used as endogenous controls due to their stable expression in our system(23).