Dear Noura
It is clearly that you have 2 bands (2 site of amplification for each sample) or you have primer dimer.
You should check by Gel Electrophoresis to know what you have then you can find your optimize conditions for your Reactions.
Good Luck
You can see that the first peak is between 75-80 degrees Celsius. If your primer melting temperature is the same, there are your dimers. If it is not the same, you have nonspecific amplification.
Are these from the same primers or two different sets? Typically, I would consider the lower Tm to be the dimers if these are in fact the same primers. However, I've found melt curves to be unreliable in detecting dimers and prefer to use electrophoresis. Since dimers are so small, a 3 or 3.5% agarose gel works much better to see them or some people go through the trouble of running an acrylamide gel. I've observed some primers to yield a pretty unreasonably high level of dimers that were not detected in the melt curve. However, if you see two peaks in the same melt curve, it usually is because of dimers.
To be able to help you we need to know:What is the expected size of your amplified product and what is the sequence of your primers and melting What is the expected size of your amplified product and what is the sequence of your primers? and melting
What is the expected size of your amplified product? and what is the sequence of your primers and melting What is the expected size of your amplified product and what is the sequence of your primers?
What is the sequence of your primers? and melting What is the expected size of your amplified product and what is the sequence of your primers?
What is melting temperature of your primers and your product?
Best way to know that is by including a negative control (-template) in your qPCR reaction. The product peak will be absent in -template reaction and thereby you can identify the peak(s) corresponding to primer dimers.
What is the expected size\Tm of your amplified product? i think you have 2 products, with 2 different Tm (75-80 and 80-85), primers dimer always have a smaller Tm than that, i think you have a non specific band, it is easy to check with electrophoresis.......good luck
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