Ligating small insert

ijiwaru at wheel.dcn.davis.ca.us ijiwaru at wheel.dcn.davis.ca.us
Tue Aug 20 13:47:40 EST 1996


Minh,

I've done a couple different clonings of the size you mention, 42bp
NsiI-BamHI into pGEM7Zf+ SmaI-BamHI and ~50bp SacI-EcoRI of pSP64-polyA
into pGEM4.  As I remember both times I used about a 20-50-fold molar
excess of insert to vector with all fragments being gel isolated.  I
didn't have too much of a problem with multiple inserts or background
either time.

Good luck,

Lyle Najita
Plant Pathology
University of California - Davis

In article <321A3780.738E at ea.oac.uci.edu>, "Minh Q. Nguyen"
<mqn1 at ea.oac.uci.edu> wrote:

> Hi,
> 
> Has anyone tried ligating a very small insert (~50 bp) to your typical 
> vector (e.g. Bluescript)?  If so, what is the optimal ratio of insert 
> to vector and how efficient is the ligation?  Any suggestion would be 
> appreciated.  Thanks.
> 
> mqn



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