Dear all,

Thanking you in advance for your attention, I would like to ask you two questions regarding an electrophoretic run on 2% agarose gel:

1) I did an electrophoretic run of the Real-time PCR products (with Sybr green). Although the band of my target product is evident, I notice some very slight sub-bands (indicated by the arrows) which, according to the molecular scale, could have a size of 500 bp. I designed the primers by myself, and making a blast they are specific for my gene of interest . What do those sub-bands represent? Can they affect the analysis of fluorescence?

2)The same primers gave me products of a different size (in the photo, the circled bands). The primer targets a gene that has 3 transcript variants. Each well has a different individual (animal). Could those two circled products be a transcript variant?

Thanks a lot for you precious help,

Best Regards

Matteo

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