Cohesive end ligation of dsDNA to ssDNA

D.K. dk at no.email.thankstospam.net
Thu Nov 29 22:13:09 EST 2001


In article <X8CN7.89$t4.2591 at news.uchicago.edu>, "Emir Khatipov" <khatipovNO at NOuchicago.edu> wrote:
>I think I have a nontrivial question here for cloning gurus.
>Would ligase (T4 or Ec, or RNA ligase) ligate the following molecules to
>each other:
>1) vector linearized with 2 different 6-cutter restrictases (4-base
>overhangs),

Basically, no. 

>2) single-strand (ss) oligo with ends complimentary to the sticky ends on
>the linearized vector?
>vectorxxx    <--tgca-random-oligo(50bp)-ttgg--> xxxvector
>vectorxxxacgt                                                aaccxxxvector

Yes. 

>So the question is does the 3-strand ligation work, and if work, how
>efficiently?

IMO, about the same as regular sticky ends. 

DK




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