promoter deletions

Dilip Dias MAD2631 at
Thu Apr 4 14:49:13 EST 1996


I have used kits from two different companies Promega and NEB Both give 
the deletions ladder you expect.  They are easy to use but for both 
these kits to work the starting DNA needs to be free of nicks. The best 
and the sure way is to cesium chloride purify the DNA to remove the 
nicked (therefore linearized plasmids). We had some difficulty in 
isolating the exact exodeleted plasmid (the one with the predicted size) 
once transformation is done and this is still true.  You may get lot of 
colonies but all of them will not have the size deletion you are looking 
for. only about 25 - 40% will have if you are lucky.  So when screening 
for colonies grow atleast 10 for each time point deletion to get atleast 
one you need. Hope this helps.

Good luck

Dilip Dias
Dept. of Forest Science
Texas A & M University

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