title: Deep sequencing of SMB-RES8 miRNA-Seq of mus musculus
design: SMB-S15 cells were exposed to 10 M resveratrol solved in DMSO  for 8 days, nominated as SMB-RES4. The collected supernatants from the cell preparations were centrifuged in order to remove cellular debris. The exosomes were extracted by using a commercial ExoQuick-Tc kit (SBI). 5 g of total RNA each sample was size-fractionated by 15% polyacrylamide electrophoresis (PAGE). The small RNA fraction (18-30 nt) was extracted and ligated to 5- and 3- RNA adaptors using T4 RNA ligase. The RNA-adaptor constructs were purified and reverse transcribed. Then the reverse transcribed products were amplified using the following PCR (Polymerase Chain Reaction) program: a 15-cycle reaction at 98C for 30 sec, followed by 15 cycles of 98C for 10 sec, 72C for 15 sec, and 72C for 10 min. Ultra-deep sequencing was performed using one flow cell channel per sample.