Mirneasy serum plasma kit 50

All the patients signed an informed consent before the neuropsychological tests, and the blood samples were collected on the day before surgery and 7 days after surgery. Meanwhile, all the blood samples were examined, and it was made sure that they were not in the hemolysis state. After having been collected by one professional registered nurse, the whole blood was left undisturbed at room temperature for about 30 min. Following centrifugation at 3,000 × g for 10 min at 4°C, the supernatant (serum) was collected. We also confirmed that the color of the serum was clear and yellowish. Then, the supernatants (serum) were transferred to a refrigerator at −80°C for further analysis. The total RNA was extracted using the miRNeasy Serum/Plasma Kit (50) (Qiagen, Germany). The concentrations of the RNA samples were determined spectrophotometrically at 260, 280, and 230 nm by using a NanoDrop ND-1000 instrument.

Total RNA was extracted from serum using the miRNeasy Serum/Plasma Kit (50) (Qiagen, Germany). Quantitative real-time PCR (qRT-PCR) was conducted with an Eppendorf RT-PCR system (Hauppauge, NY, United States). We used Caenorhabditis elegans-miRNA 39 mimic (Ce-miR-39) as miRNeasy Serum Spike-In control. The specific primer pairs used in the study are listed in Table 1.