I am trying to sort out this primer dimer (in NTC) issue for a very long time. After many of my previous trials in which NTC and my sample bands looked same, only in this trial the NTC and my sample bands look different upon using DMSO and Betaine. My target size is 196 bp. Has anyone got these kinds of results before or can anyone kindly help me interpret it?. I have attached the image along with the labeling indicating the amount of DMSO and Betaine used. we have HMB based identification kit in our lab, but I think it would be pointless if my NTC showed bands too. Do I have to change my primers
Hi,
In general, the maximum DMSO concentration used in PCR assay is 10%. But, your experiment was performed with much higher concentration (14-16%). Likewise, the optimal betaine concentration is 1.0mM.
Please try with the optimal concentration: DMSO (6%) and Betaine (1mM) often works well.
Follow this link to have better understanding on pcr additives in lamp pcr
https://www.nature.com/articles/srep37837#f2
All the best
Dr.Kalyanaraman
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