nested deletions

R. Woodward rw200 at
Fri Aug 18 11:07:30 EST 1995

Hello everyone
 Ineed a bit of advice re: nested deletions. I have been using a kit supplied by NEB ('Exo Size deletion kit') the problem is that the only plasmid I can rescue are of parental size evne though I can see that the ExoIII has worked very nicely from analytical gels of samples taken at different time points of my deletion reactions. I suspect the Mung Bean Nuclease is to blame (the reaction vol is 20ul for 30min at 30oC using 50mMNaOAc pH 5.0, 30mM NaCl and 1mM ZnSO4). Is there anything I should be aware of a
bout MBN any suggestions etc.... I have used the Pharmacia deletion kit in the past and that worked o.k. but that used S1 nuclease. Whats the feeling should I buy some S1 nuclease to use in my present kit?  Any other thoughts would be welcomed. Thanks v.much to all of those poeple who have answered my previons questions.

 Email rw200 at
 Dept Pharmacology
 Tennis Court Rd
 University of Cambridge
vi -r /usr/tmp/nn.a04526

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