Baffling cloning results
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:
> -- 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).
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
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