GSM5135521: ICH9-A; Homo sapiens; RNA-Seq
The total RNA of peri-hematomal tissues from 11 patient with ICH was extracted using TRIzol reagent (Invitrogen) following the manufacturer's protocol and RNA integrity was measured. Magnetic beads with Oligo (dT) are used to isolate mRNA (for eukaryotes) or by removing rRNAs from the total RNA (for prokaryotes). 1-5 μg of total RNA was removed ribosomal RNA. The quality of RNA (ribosomal RNA 28s:18s > 1.7) for brain tissue was performed the further transcriptome analysis. Mixed with the fragmentation buffer, the mRNA is fragmented into short fragments. cDNA is synthesized using the mRNA fragments as templates. Strand-specific RNA-seq data was generated. During the QC steps, Agilent 2100 Bioanalyzer and Applied Biosystems StepOnePlus Real-Time PCR System are used in quantification and qualification of the sample library. Library preparation was established using the Illumina TruSeq Stranded kit and sequenced on an Illumina Hiseq 4000 sequencer. Libraries were barcoded and sequenced in one lane, thus avoiding lane effects, to obtain 100 bp paired end reads. An average of 30 million reads were obtained for each sample.