GSM6600251: platelet treated with SARS-CoV-2 E protein, rep2; Homo sapiens; RNA-Seq
Fresh blood collected was subjected to platelet isolation within 1 hours after blood collection. Platelets were incubated with SARS-CoV-2 E protein or PBS for 1 hour at 37℃. Then the platelet pellets were isolated by two continuous steps of centrifugation, followed by lysis in QIAzol (QIAGEN) and extracted the total RNA using the miRNeasy Mini Kit (QIAGEN). Total RNA was qualified and quantified using a Nano Drop and Agilent 2100 Bioanalyzer (Thermo Fisher Scientific, MA, USA). Totol RNA of platelets was isolated by using the miRNeasy Mini Kit (QIAGEN). Platelet RNA was eluated in 30 μL elution buffer.Total RNA was qualified and quantified using a Nano Drop and Agilent 2100 Bioanalyzer (Thermo Fisher Scientific, MA, USA), and included as a quality standard for subsequent experiments only platelet RNA samples with a RIN-value >7 and/or distinctive rRNA curves. The double stranded PCR product from previous step was heat denatured and circularized by the splint oligo sequence. The single strand circle DNA (ssCir DNA) was formatted as the final library. The final library was amplified with phi29(Thermo Fisher Scientific, MA, USA) to make DNA nanoball (DNB) which had more than 300 copies of one molecular, DNBs were loaded into the patterned nanoarray and single end 50 bases reads were generated on BGISEQ500 platform (BGI-Shenzhen, China).