I would suggest taking a look at the lysis conditions you are using.
For example, are you using only lysozyme or lysozyme plus mutanolysin
(the latter is better)? Are you adding the enzyme(s) to the cell
suspension/agarose mixture before casting the plugs or to the lysis
buffer after the plugs are cast? I would be glad to give you some
specific comments depending on what you're doing now.
Dept. Med. Microbiol. and Immunol.
Creighton Univ. Sch. Med.
M. DAUGHERTY wrote:
>> Have recently started doing pulsed field gel electrophoresis on
> vancomycin-resistant enterococci, using standard techniques. Does
> anyone have suggestions for improving the quality of the runs? The
> leading bands are blurred, and reproducibility is not great. Thanks for
> any thoughts.