disease state: HACI tissue: Plasma gender: male age: 73 clinical outcome (mrs on discharge): 5 stroke subtype (toast): cardioembolic infarct stroke subtype (ocsp): total anterior circulation infarction
Treatment protocol
Blood samples from the HACI group and the HVT group were obtained and immediately centrifuged. The supernatants were transferred to RNase-free tubes and stored at -80°C.
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted and purified using mirVanaTM PARISTM(Cat # AM1556, Ambion, Austin, TX, US), following the manufacturer’s instructions and checked for a RIN number to inspect RNA integration by an Agilent Bioanalyzer 2100 (Agilent technologies, Santa Clara, CA, US).Total RNA was quantified using a NanoDrop-2000 spectrophotometer.
Label
Cy3
Label protocol
miRNA molecular in total RNA was labeled by miRNA Complete Labeling and Hyb Kit (Cat # 5190-0456, Agilent technologies, Santa Clara, CA, US) followed the manufacturer’s instructions, labeling section.
Hybridization protocol
Each slide was hybridized with 100ng Cy3-labeled RNA using miRNA Complete Labeling and Hyb Kit (Cat # 5190-0456, Agilent technologies, Santa Clara, CA, US) in hybridization Oven (Cat # G2545A, Agilent technologies, Santa Clara, CA, US) at 55°C，20 rpm for 20 hours according to the manufacturer’s instructions, hybridization section. After hybridization, slides were washed in staining dishes (Cat # 121, Thermo Shandon, Waltham, MA, US) with Gene Expression Wash Buffer Kit (Cat #5188-5327, Agilent technologies, Santa Clara, CA, US).
Scan protocol
Slides were scanned by Agilent Microarray Scanner (Cat # G2565CA, Agilent technologies, Santa Clara, CA, US) and Feature Extraction software 10.7 (Agilent technologies, Santa Clara, CA, US) with default settings.
Description
microRNA expression in human plasma of patients with hyperacute cerebral infarction with stroke onset in 6 hours
Data processing
Raw data were normalized by Quantile algorithm, Gene Spring Software 12.6 (Agilent technologies, Santa Clara, CA, US). When the difference of one microRNA in certain sample between the signal detected by probe and background signal was not significant, it would be marked with “absent” and given a identical value, such as “-3.1303349”.
