pcr amplification
 

i have already amplified and got product one month ago. But now while doing the same sample iam not getting in the same machine. WHAT CAN BE THE REASON FOR THAT?.

Re: pcr amplification
 

Sounds like it could be a sample issue. What temperature was the template stored it? Was it stored in sterile buffer? Are your Master Mix ingredients still good? Make sure your using the right concentrations of materials, as well as correct forward and reverse primers. Also try annealing step at 55 C.