PCR Optimization for Beginners: A Step by Step Guide

Background: The invention of Polymerase Chain Reaction (PCR) marked a breakthrough in biomedical research. Its divided the timeline into an era before and after PCR. Because its multiple applications, it has become vital tool for clinical researchers diagnosticians. A effective PCR experiment requires adequate knowledge each reaction component stepby- step procedure to attain optimized results. This research aims employ optimization strategies that are easy perform, cost-effective, do not require kits generation amplicons TYR, MITF, SOX10 genes can be used sequence analysis. Materials Methods: Whole blood samples were extract genomic DNA with inorganic method. quantification was done by spectrophotometry Optimization adopted generate products candidate visualized agarose gel electrophoresis. Sanger sequencing performed check quality specificity generated via strategies. Results: study demonstrates novel approach troubleshooting failed reactions without use kits. result indicated sterilized material reagents along optimum MgCl2 concentration (2.0-3.0 mM), quantity (25 ng/μL), annealing temperature (54-60°C) necessary achieve successful amplification. Sequence analysis revealed no background noise often associated Conclusion: Optimizing concentration, quantity, temperature, contamination-free essential steps optimization. Following this guide, anyone lacking proper supervision, little or should desired