2 replies to this topic

[Danny Chow]

Does anybody using EDTA to lysate bacterial cell ?

Could you please provide me the lysate buffer ?

i am using 2 buffers

1) Tris/Cl 20mM, EDTA 2.5mM, sucrose 20% (30mins on ice) 10ml for 1L bacterial culture

2) Tris/Cl 20mM, EDTA 2.5mM (30 mins on ice) 10ml for 1L bacterial culture

I will discard the (1) solution, and then centrifuge down the pellet, then add (2)
I found that almost 50% of the target protein was appeared in the (1)

Thanks

[aimikins]

what kind of bacteria?

[Danny Chow]

aimikins, on Jan 5 2010, 07:31 AM, said:

what kind of bacteria?

E.Coli