Hello everyone

 

I am going to attach an SiRNA to a functionalized biotin.

 

Then i need a way to quantify the efficiency of the reaction and i tough that a fluorescet Streptavidin could be usefull

 

 

The problem is that i don't know exactly how to run the essay

 

I tought that i can attach the SiRNA/Biotin somehwere and then reveal it by streptavidin

 

Any suggestion?

 

thanks a lot

 

Francesco

Without much thinking I would simply try a more standard approach and use streptavidin to extract biotinylated siRNA (i.e. performing affinity purification). The siRNA yield can easily be quantified and higher yield indicates higher biotinylation.

Thanks for your answer

 

do you think that using streptavidin conjugated with a resin or beads should be ok?

 

then i can elute... and how i can quantify the Biotin-SiRNA conjugate?

 

Francesco

Actually, scratch that. The easiest way would probably run biotinylated vs non-biotinylated samples in a gel. Or add streptavidin for stronger separation (due to larger size differences).