I need to measure the expression of special variant of a gene in mRNA level, but it's not possible to design specific primers for the variant without detecting the other variants. So can anyone please suggest some methods for this purpose. I don't want to use probes.
Ryan M Boileau
How is it a variant of the original gene? On the transcript level it still seems you would be able to design internal primers to detect alternative splicing if that was the variance described.
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Jill Muhling
How exactly does the variant differ from the original gene? If you're reverse transcribing into cDNA then try melt curve analysis to look for differences in melting temperature
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Silvia Franzellitti
what about using a Taqman assay with the same primer pair for both variants but variant-specific Taqman probes?
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