Any answer for both?

I suggest you use lysostaphin for

staphylococcus aureus.

Regards

Hello dear! The protocol you have used seems ok.

But one thing you didn't mention, have you been able to concentrate the plasmid DNA by cleaning the solution?

For e.g: By Phenol or Chloroform Extraction followed by Ethanol precipitation?

Dear Rajat

did you tried to extract those plasmid from s.aures and p.aeruginosa or from e.coli?

Because tiye protocol seems for e.coli and if p.aerugnosa is a gram negative not so different from e.coli, s.aureus is quite different in terms of membrane stability.

First of all, also for gram negative as e.coli i suggest ti you to use commercial kit that today are quite cheap and works very well.

give a look to the ProteoCool n°12 at Page6 of my blog: https://proteocool.blogspot.com/?m=1

moreover the first things to consider is how you can extract the dna from the cells efficiently.

For example for s.aureus preliminary incubation. with lysostaphin (attached you find a protocol suggested by qiagen) is essential to broke the cells and extract the plasmid from inside.

good luck

Manuele