I have had this problem before and the cause was that I used the same primer twice (reverse+reverse instead of reverse+fordward), but now I am pretty sure I used the correct primers. The protocol is ok and the reagents are good too.
I attached a picture of the problem. It is a qPCR with probe.
Laurence Stuart Dawkins-Hall
You need to be much more specific with your question and perhaps provide a picture of these curved peaks
Are you referring to kinks in bands on agarose gels?
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Sandra Barroso-Arévalo
I am sorry. You are rigth.
It is a real time PCR. I have attached a picture and edited the first comment. Thanks.
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Laurence Stuart Dawkins-Hall
- Your problem is low fluorescence
- What are the efficiency values for your primers?
- Did you use > 1ug of RNA to make your cDNA
- Have you checked your primers by standard RT PCR to make sure you get strong bands on a gel
- What about amplification of a housekeeper like b actin or gapdh from your cDNA: If this is weak then your cDNA is of low quality
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