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A new approach to touch down method using betaine as co-solvent for increased specificity and intensity of GC rich
Daliparthy D Pratyush1, Shalbha Tiwari, Ashok Kumar
1Department of Endocrinology and Metabolism, Banaras Hindu University, Uttar Pradesh, India.
Abstract:
Tissue specific genes that contain high GC segments are difficult to amplify by standard PCR. We report an improved method for successful amplification of gene segment that has >70% GC base pairs. This new method of touch down PCR differed by having an initial annealing temperature (Ta) 1.5°C below the primers melting temperature that descended 0.2°C per cycle for 20 cycles and continued thereafter at fixed Ta for next 15 cycles. Different co-solvents were tested with this method to improve the result and betaine proved better than the other co-solvents. This new method is economical, fast and specific in amplifying GC rich region of other genes also.
