Optimization of proteinase K incubation protocol duration during DNA extraction from oral squamous cell carcinoma FFPE samples

Background: Formalin-fixed paraffin-embedded (FFPE) specimen archives are a valuable source of sample material for molecular biological analysis. However, the DNA isolated from FFPE samples is usually low in concentration and fragmented. Thus, it necessary to optimize extraction protocol obtain best results. Proteinase K incubation undoubtedly crucial procedures, but this step often not well explained manufacturer’s manual. Purpose: This study aimed find optimal duration proteinase protocols achieve highest yields. Methods: Fifteen paraffin blocks Oral Squamous Cell Carcinoma (OSCC) specimens were obtained, cancerous areas microdissected into smaller cuts extraction. The randomly divided three groups (n=5) subjected different protocols: one-hour at 56ºC as per instructions (Group I), 24-hour II), 48 hours room temperature with an additional four III). extracted was then quantified using Nanodrop spectrophotometer. recorded data analyzed ANOVA-LSD. Results: found Group III (107.74 ± 41.92), which significantly higher compared II (59.46 30.32) I (6.46 1.97) (p<0.05). Conclusion: In conclusion, modifying can lead yield study, most optimized incubation, resulting yields, 56ºC.