Deletion of plasmid

Dr. Duncan Clark duncan at nospam.demon.co.uk
Wed Jul 15 04:26:28 EST 1998


In article <35AC33BA.2B0A at bioneer.kaist.ac.kr>, Jung Hoon Ahn
<jhahn at bioneer.kaist.ac.kr> writes
>But strangely, after ligating two, transfoming it into E.coli XL1-Blue,
>mini-prepared plasmid is not what I expected. The front part of gene is
>deleted.
>I inserted the rest part of gene into the cloned gene in pUC19.
>What is the reason of this phenomenon and how can I solve this probelm.

If the gene is expressing and lethal to E.coli then the E.coli will
either die or the gene will be deleted. Try cloning opposite to the lac
promoter, use pBR322 or a lower copy vector, don't clone it with a
promoter etc.

Duncan
-- 
The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

Duncan Clark
DNAmp Ltd.
Tel: +44(0)1252376288
FAX: +44(0)8701640382
http://www.dnamp.com
http://www.genesys.demon.co.uk



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