Breaking E. faecalis

[Leigh Schmidtke]

In article <322C889D.468E at bmb-fs1.biochem.okstate.edu>, Ron Tate
<rtate at bmb-fs1.biochem.okstate.edu> wrote:

> Does anybody have a consistant method for breaking E. faecalis?  I
> typically use sonication but for me it is horribly inconsistant and
> takes quite a while, I would appreciate hearing your experiences.
> -- 
> >>>>>>--------------------------->
> Ron
> Tate                                                                   
> Lab of Franklin Leach
> Dept. of Biochem. & Molecular Biology
> Oklahoma State University 
> rtate at bmb-fs1.biochem.okstate.edu
> (405) 744-9326
> ---------------------------------------------

You could try an incubation step with lysosyme (450 000 U/mL, 60 minutes
at 37šC) before sonication.  I found this worked well for a range of
Enterococci/Streptococci and Lactococci.  If you sonicate, make sure you
place your sample on ice.  I only used short bursts of say 15 seconds to
prevent boiling the sample (sample volumes were <0.5 mL).  Depending on
what you want to get from the bugs, you could also solubilise with SDS
after lysosyme treatment.  For a complete run down of the method see;
Schmidtke and Carson. Journal of Applied Bacteriology. 1994. 77:229-236.
Hope this helps.

Leigh Schmidtke
Department of Biomedical Science
University of Tasmania
Leigh.Schmidtke at appbio.utas.edu.au