Protocol Online logo
Top : New Forum Archives (2009-): : Molecular Biology

Can "3' - biotinylated oligo" be ligased with "5' phosphoryl - (Dec/22/2012 )

I have 3' biotinylated oligo, Can it be ligated to 5' end of oligo? On what site of DNA a biotin linker is covalently connected?


A 3' biotin will block ligation. But a 3' biotin-dT base could potentially be ligated. I don't know how efficient this would be, or if standard ligases would work.


3'-biotin will block OH thats why ligation is not possible. here are some links, which might be helpful-


phage how is biotin-dT base different? ..


Biotin-dT links the biotin to the thymine base, rather than to the 3' -OH group on the ribose. This leaves the 3' -OH available for linkage to the next base. Steric issues may make ligase not work or be inefficient, but the block is no longer an issue.