GSM2358306: MELAS with m.A3243G mutation [sequencing]; Homo sapiens; ncRNA-Seq
Total RNA was extracted using Trizol Reagent (Invitrogen, CA, USA) following the manufacturer's procedure. The total RNA quantity and purity were analyzed using Bioanalyzer 2100 and RNA 6000 Nano LabChip Kit (Agilent, CA, USA) with RIN number >8.0. Approximately 5 ug of total RNA was used to deplete ribosomal RNA according to the manufacturer's instructions of the Ribo-Zero rRNA Removal kit (Epidemiology version) (Epicentre, an Illumina company, Madison, WI). Following purification, the RNA is fragmented into small pieces using divalent cations under elevated temperature. Then the cleaved RNA fragments were reverse-transcribed to create the final cDNA library in accordance with the protocol for the RNA-Seq sample preparation kit (Illumina, San Diego, USA). The average insert size for the single-end libraries was 50 bp. We then performed the single-end sequencing (50 bp) on an Illumina HiSeq 2500 at the LC-BIO (Hangzhou, China) following the vendor's recommended protocol.
GEO Accession
GSM2358306