Dear Muhammad

Because it may be disabling the taq DNA polymerase due to successive cycles, it is better to run first.

if these are dna samples where you have very littledna then I think that the buffer,Mg , primers and dna will be fine so I would add dNTPs with more enzyme and expect the pcr to work. otherwise just run it properly as it should have happened first time

Okay i will run it again and let you know thank you 😊