title: miRNA-Seq of MC3T3 cell Growth condition replicate 2
design: Isolation of exosomal miRNA samples for next generation sequencing (NGS) were processed and used for NGS by System Biosciences (SBI). 10 ml of cell culture media was centrifuged at 1500 x g for 5 min to remove residual cells and debris. The supernatant was transferred to a new 50 ml conical tube for exosome isolation. NGS libraries were generated with the TailorMix Micro RNA Sample Preparation version 2 protocol (SeqMatic LLC, Fremont, CA). Briefly, 3'-adapter was ligated to the RNA sample and excess 3'-adapters were removed subsequently. 5'-adapter was then ligated to the 3'-adapter ligated samples, followed by first strand cDNA synthesis. cDNA library was amplified and barcoded via enrichment PCR. Final RNA library was size-selected on an 8% TBE polyacrylamide gel.