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Can "3' - biotinylated oligo" be ligased with "5' phosphoryl

ligation biotin

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3 replies to this topic

#1 lioiviei14

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Posted 22 December 2012 - 02:36 AM

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?

#2 phage434

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Posted 22 December 2012 - 11:52 AM

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 neuron

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Posted 26 December 2012 - 02:53 AM

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

https://eu.idtdna.co...gold-particles-

http://www.piercenet...?fldID=06010705

phage how is biotin-dT base different? Posted Image ..

Edited by neuron, 26 December 2012 - 02:56 AM.


#4 phage434

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Posted 26 December 2012 - 05:09 PM

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.





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