With hot start kit, it pcreasily amplify your fragments. You have to check your primers once again. For obtaining good high yield, parameters like initial template DNA, MgCl2, dNTPs and extension time have to be optimize. Do pcr with gradient temperature with +/- 5 of Tm value of primer extend PCR cycle from 28 to 35 cycle. sterilize all materials for gel electrophoresis and confirm presence of Raw DNA/ Primer/ dNTPs, if these present there is a clear problem in your PCR so do optimization.

Thanks for your attention.

Hi Maryam

Don't worry about not getting amplification...something must have gone wrong....If you are doing PCR for the first time..take help of somebody in the lab/department who is doing it regularly and do it under hos guidance....In case you are doing it regulalry but this time there is some problem.....use a positive control which has always worked in your hands...This will give you indication about the possible problem.....

Thanks sir

I don't know any a bout it please explain me how to do it?