In human tumors, somatic mutation frequently occur in K-ras gene at codon 12, which makes the K-ras protein hyper active leading to uncontrolled signaling for cell division: one of the important hall mark of cancer. In order to correlate mutations in K-ras to cause, response to treatment, disease progression and recurrence of Head and Neck Squamous Cell Carcinoma (HNSCC) the following study was undertaken. By using PCR–RFLP method prevalence of codon 12 in K-ras gene was studied in 56 HNSCC patients. High frequency of K-ras mutation was detected in codon 12 (60.71%). The result of this study helps us in understanding the role of K-ras somatic mutations in HNSCC patients and in designing novel treatment protocols for HNSCC patients. © 2020, Association of Clinical Biochemists of India.