Im new to qpcr field. my question is whether the dead samples or live samples is better for qpcr? i am doing qpcr on fish n crustacean ectoparasites, and both are clearly dead. for fish, as i use universal primer i can get a result, so i guess it is okay to use dead samples but unfortunately for my ectoparasites, i couldnt get any result using the primer that i design from crab n shrimp, after all optimization. I design primer from crab and shrimp as i there's no gene expression done in any parasites for gene that i want.
From my experience with mouse samples (without parasites to my knowledge) I can only recommend to preserve the samples as soon as possible, RNA degrades very fast. Its also a question of the abundance of your RNA of interest, if you look for house keeping genes the chances will be probably higher.
Hi.
I agree with Mr. Hoffmann. Did you checked your primer using genomic DNA as template? If yes and it is not working I would design new primer.
Hi
If dead specimens has been freshly preserved , they are ok, it means that they immediately were put in a good preservative.
But, if they have been left away and there has been lysis of tissue, then you will not have enough cells.
Regards
Abdigoudarzi
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