GSM2264263: Stroke patient 31; Homo sapiens; miRNA-Seq
Total RNA isolation was performed by Seqmatic LLC (Fremont, CA). Briefly, TRIzol reagent (Thermo Fisher) was used to extract RNA from the cell samples Libraries were generated using total RNA prepared by SeqMatic. The SeqMatic Tailormix miRNA library prep kit V2 was used to create libraries that are compatible with Illumina sequencing. Each sample was validated for yield and quality using a BioAnalyzer High Sensitivity DNA assay. Libraries were normalized and pooled in equal molar ratios. The pool was gel purified and the purified product was quantified using the SeqMatic TailorMix HT1 qPCR assay. A single end read 50bp sequencing run was performed using the Illumina MiSeq instrument. Reads were demultiplexed by barcode and output into FASTQ format for subsequent analysis. Raw sequence data was trimmed of adapters and low quality sequences using Trimmomatic. Only reads with a minimum Qscore of 15 and length of 16nt were kept for downstream processing.