siRNA Ligation problem
(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
> i gave top strand sequence in 5' to 3' direction and bottom strand
> in 3' to 5' direction. However the bottom strand has been sequenced and
> 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'
> of current oligo (that will be desired 3' end of bottom strand)?
> i hope my description of problem is clear.
> need help
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