GSM5600192: TR4333; Homo sapiens; RNA-Seq
GSM5600192
Blood collected in Gdańsk was processed within 48 hours upon collection, strictly according to thromboSeq protocol [PMID: 30894694]. Briefly, tubes were centrifuged to obtain platelet rich plasma (20 minutes, 120g), pelleted (20 minutes 360g) and carefully resuspended in RNAlater (Thermo Scientific). After overnight incubation at 4°C, platelets were stored at -80°C. Blood collected in Gdańsk was shipped to Cancer Center Amsterdam, VU University Medical Center for further processing. RNA was extracted with mirVana miRNA isolation kit (Ambion, Thermo Scientific). For RNA quality verification the RNA 6000 Picochip (Bioanalyzer 2100, Agilent) was used. Subsequently, SMARTer Ultra Low RNA Kit for Illumina Sequencing v3 (Clontech) was used for cDNA amplification. Next, cDNA was subjected to nucleic acid shearing by sonication (Covaris Inc) and then labeled with single index barcodes for Illumina sequencing using the Truseq Nano DNA Sample Prep Kit (Illumina). Labeled platelet cDNA library was measured with the DNA 7500 chip or DNA High Sensitivity chip (Agilent). High quality samples were pooled and submitted for 100 bp Single-Read sequencing on the Illumina HiSeq 2500 and HiSeq 4000 platform. 100 bp single-end reads. Standard Illumina protocol