Real time quantitative PCR analysis of serum circulating miRNAs from patients with head and neck tumors. [exp1]

The samples were drawn from patients with diagnosed oropharyngeal cancer treated with irradiation. We used blood samples (10ml) collected from patients 7 days after receiving 20Gy dose of irradiation. Blood was centrifuged for 10 min, after which serum was aspirated, aliquoted in RNAse-free tubes, stored at −80°C until required for further processing and did not undergo more than two freeze-thaw cycles. MicroRNA was extracted from 200 μL serum using miRCURY RNA Isolation kit (Qiagen) following the manufacturer’s protocol and stored at −80°C. miRNA qPCR assays from 20 patients from after 20Gy irradiation due to oropharyngeal cancer. Equal amount of RNA was used from each patient.