DNAses/Nucleases inhibitors

Vivian Miao vmiao at oregon.uoregon.edu
Thu Feb 25 17:39:11 EST 1993


In article <1993Feb25.171337.26865 at gserv1.dl.ac.uk>, N.SALAZAR at lshtm.ac.uk
(Nelson Salazar) wrote:
> 

> Does anybody have any experience using Auryntricarboxylic Acid-
> Ammonium salt (Aluminon) as a DNAse inhibitor??? Would this help to
> avoid smearing (degradation) of chromosomal DNA preparations when
> preparing the agarose blocks ??? I am trying to obtain chromosomal DNA
> of Entamoebas but after running the CHEF gel I get a smear of DNA.
> Perhaps the voltage ??? I have used 160 volts..


ATA didn't make any difference to my chromosomal plug preps,
 although I didn't have "a lot" of smearing to begin with
 (if you want more details, email me).  In addition to the
 suggestions from NEB, perhaps you could test whether the
 smearing you're getting is really due to nucleases by adding
 some known DNAs to one of your samples when you next make
 them, and then asking if that DNA is degraded?  Known DNAs
 could be just some plasmid (run under regular electrophoretic
 conditions - you just want to know if it survives
 co-incubation with the chromosome preps) or lamda, etc. 

Vivian

(vmiao at oregon.uoregon.edu) 



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