Hi everybody,
I'm currently working on circulating miRNAs.
I performed miRNA microarray and found some differentially expressed miRNAs between patients and healthy subjects.
I have now to perform differential expression validation using RT-PCR.
I chose the poly-T adaptor PCR method (on which many commercial kits are based) and I was looking for an exogenous control to use.
I see many using cel-miR-39 in the RNA isolation stage, but I already have total RNA so I have to add it as a spike-in in my polyadenylation mix.
I would like to know if anyone has a protocol or at least knows the correct concentration of cel-miR-39 to spike in at the polyadenylation /cDNA synthesis stage.
Thank you so much ,
Any help would be really appreciated.
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- Biological Science
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