RNA-Seq of human Oral squamous cell carcinoma-derived exosomes: normoxia 2
Total RNAs from the normoxic exosomes was used for miRNA library preparation and sequencing. Total RNA samples were fractionated on a 15% Tris-borate-EDTA (TBE) polyacrylamide gel (Invitrogen) and small RNAs ranging between 18 and 30 nucleotides (nt) were used for library preparation using the Illumina TruSeq Small RNA Sample Preparation Kit. After ligation of 3ĄŻ and 5ĄŻ adapters, small RNAs were reverse transcribed and amplified by PCR with primers complementary to the adaptor sequences. Finally the amplified PCR products obtained after gel purification and quality control were sequenced using the Illumina HiSeqTM 2500 platform.