I am trying to find a way to functionalize purified genomic DNA.
That is, I need a way to add reactive handles (amines, carboxylic acids... any conventional chemical group, really) on a linear, natural DNA, with an efficiency high enough to graft it to activated surfaces, incorporate it into polymers etc. So, something like 1 reactive handle every 102-103 bp would be a minimum. The downstream applications are chemistry rather than molecular biology, so I am interested in reactions that would be applicable to fairly concentrated samples (above 1mg/mL) and large volumes, which rules out enzymatic reactions.
I have tried to use the primary amines of bases (A & G) for reaction with epoxides or aldehydes, but I could not access these despite DNA denaturation. I also considered shearing the DNA to access reactive terminal phosphates, for EDC/Imidazole functionalization, but cutting the DNA is detrimental for downstream applications.
On a side note, because the DNA is fairly concentrated, reactions cannot be performed in acidic conditions because DNA would precipitate.
Any ideas? Thanks!
I do not think you can do this kind of functionalizition effficiently by pure chemistry, you need to do it by enzyme biologically. Those NH2 groups on A G C are not really primary amines because they are not on saturated alkane, they are not very reactive, they are like NH2 on aniline. You need much harsh condition for those NH2 groups to react with epoxides or aldehydes , even so, it will not very efficient.
But you can introduce H2NCH2CH=CH-dU on the end of DNA by enzyme with H2NCH2CH=CH-dUTP, I do not know how, I am not a molecular biologist, I only know it can be done this way.
Good look!
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