Baffling cloning results

Ashok Aiyar aiyar at ebv.mimnet.northwestern.edu
Wed Aug 29 09:00:10 EST 2001


On 29 Aug 2001 13:34:06 GMT,
    Karthik Aghoram (kaghoram at unity.ncsu.edu) wrote:
> Briefly:
> 
> -- both insert and vector are cut with SacI and SmaI.
> -- Bands are eluted with Qiagen's QIAEX kit.
> -- Ligated with excess insert (~3:1).
> -- SURE cells transformed.
> -- Plated on LB-Kan (50 ug/ml Kan) plates.
> 
> Result:  LAWN!

umuC is insertionally inactivated by Tn5 in SURE cells (and SURE 2).
They are therefore kanamycin resistant.

Change the resistance marker in your vector to anything other than
Kan-resistance or Tet-resistance (SURE cells also carry an F' with Tet).

Later,
Ashok
-- 
Ashok Aiyar, Ph.D.
Assistant Professor                    email: a-aiyar at northwestern.edu
Department of Microbiology-Immunology           office: (312) 503-2524
303 E. Chicago Avenue, WARD 4-123                  lab: (312) 503-2542
Northwestern University, Chicago, IL 60611         fax: (312) 503-1339




More information about the Methods mailing list