I have checked annealing temperature through gradient PCR.
I have checked my primer in NCBI.
I have checked PCR reagents concentration.
Please suggest me anyone
thanks
You probably have DNA contamination You can solve the problem by reducing the concentration of primer and increasing the anling temperature
The best technique for such cases is to use the Touch-down PCR method
Thank u Sapna and Heidar for your suggestion i will check.
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