Except the explanation, the annealing temperature may be too low for your PCR.

You can increase the annealing temperature 1◦C per trial. Or it might be Increased concentration of Primer or template.

If your target gene band size is not exist in the non-specific banding pattern, your have to review the specificity of your primers.

Best luck for you boss

Hi Shami,

There are several ways to increase specificity of a PCR assay.  I agree that increasing the annealing temperature is a great place to start.  If this is an assay that you will be repeating multiple times, I would suggest that you run it on a gradient cycler to optimize the assay, so that you maximize sensitivity while simultaneously maintaining specificity. Other options include redesigning primers or using a real-time format with a probe to add specificity.