siRNA Ligation problem

ChenHA via methods%40net.bio.net (by hzhen from freeuk.com)
Thu Jun 28 23:25:01 EST 2007


Sudheendra Rao N R wrote:
> Hi all,
> i wanted help in a T4 DNA Ligation reaction. While ordering the siRNA 
> oligos
> i gave  top strand sequence in 5' to 3' direction and bottom strand 
> sequence
> in 3' to 5' direction. However the bottom strand has been sequenced and 
> sent
> to me in 5' to 3' direction. 

Doesn't that mean you got the wrong sequence then?

(Do they allow you to order oligo in 3' to 5'?)

> Hence there is no free OH group at 3' end of
> bottom strand (which is the 5' end of sequenced oligo)
> and because of it my ligation reaction is not working. Is there any step
> which will help me subject the bottom strand and get free OH group at 5' 
> end
> of current oligo (that will be desired 3' end of bottom strand)?
> 
> i hope my description of problem is clear.
> 
> need help
> 
> regards
> sudheendra.
> 


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