I have several cells transfected with a differents gene deletions fused to GFP and in order to make a qPCR to detect all the constructs, not only for the specific gene, I tried a commercial GFP marker assay from fisher (Mr03989638_mr) ,but unfortunately it did not hybridize with any of my samples, all my constructs were on LVX-acGFP1 from Takara (Clontech). Although the nucleotide sequence have little differences , protein has >90% identity, I do not understand why the assay it is not working.
I would appreciate any suggestion.
Yoav Lubelsky
eGFP and AcGFP are different enough that a lot of tool designed for one, such as antibodies and primers, will not cross-react. You'll have to find, or design yourself. a qPCR assay for AcGFP.
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