Dwindling Plasmid yield *HELP*

Martin Leach leach at mbcrr.harvard.edu
Wed Apr 20 06:40:12 EST 1994

Dear netters,

Ok, here's the situation. I've got a couple of important deletion
constructs (containing a piece of a promoter) in luciferase reporter
constructs (promega).

In the promoter-luciferase constructs (no SV40 enhancers) i am finding
several constructs have very little yield after maxiprepping. I use the
Qiagen tip method - (i know it is not the columns or solutions as other
plasmid preps come out fine while some consistently come out crap!

I have tried re-transforming the bad plasmids into bacteria, but so far no
joy. Someone else in the lab is having a problem with this
promoter-luciferase construct also (different promoter though).

If anyone can suggest anything that may help I would greatly appreciate it.



p.s. I am gonna try terrific broth vs. Luria Broth (we usually use this), i
am also going to increase ampicillin from 25ug/ml to 100ug/ml.


.....          Martin Leach                Email:leach at mbcrr.harvard.edu 
   _|____      Dept. of Pharmacology       Phone: (617) 638-5323        
   / o  /      Boston Univ. School of Med. Fax:   (617) 638-4329         
 _/  |-/__==/  80 E. Concord St. (L603)
(BULLDOZER) \_ Boston MA 02118            "Not the old underpants on your
               USA                           head.....WIBBLE" -BLACKADDER  

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