Biotin-Streptavidin question!

[Cesco_NZ]

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

[CharonY]

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.

[Cesco_NZ]

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

[CharonY]

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).