Blunt end ligation debate. >>GEL PURIFY YOUR VECTOR

[BENNY SHOMER 9238]

Dennis J. Templeton (djt2 at po.CWRU.Edu) wrote:

: In a previous article, JANIEMI at FINABO.ABO.FI (Jarmo Niemi) says:

<Stuff Deleted>

: Here's my 2cents

: Many people who tell me that their cip doesn't work say that because their
: cipped vector prep gave "background" colonies.  In almost all of these
: cases the complainant did not gel purify their vector.  Given that, they
: cannot be sure if the failed step was the cip, or an incomplete digest of
: the starting vector....

True. Unless you (as I always do...) include a control in your transformation
assay, of restricted/purified UNligated vector. If your vector is PURE linear,
you should get here practicaly zero colonies.


: dennis

Benny.



 **************************************************************
* Benny Shomer                                                 *
* Tel-Aviv University                                          *
* Sackler School of Medicine, Dept.of Embryology and Teratology*
*--------------------------------------------------------------*
* Snail:  Ramat-Aviv , Tel-Aviv  69978 ,  Israel.              *
* E-mail :  pc386 at ccsg.tau.ac.il                               *
* Tel :  972-3-640-9238     FAX :   972-3-642-2046             *
*                                                              *
 %%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% 
%        So Many Computers , So Little Time ...                %
%                                                              %
 %%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%