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Brucella LPS extraction


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#1 saz9961

saz9961

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Posted 21 September 2004 - 04:24 AM

I'm working on a bunch of projects, detecting various bacteria using Surface Plasmon Resonance.

I'm after a simple protocol to obtain Brucella LPS. Just really looking for a crude prep that will contain relatively high amounts of LPS, not for use as an immunogen. I'm aware of the hot-phenol extraction and cold butanol protocols, but [1] I don't fancy carrying out a hot phenol extraction; our lab has limited chemistry facilities [2] We don't have ultracentrifugation facilities [3] Our source of Brucella abortus supplied formalin or heat killed cells at a concentration of 20-50mg/ml, at a cost of around $70 for 5g wet weight. All the protocols I have read assume a large fermenter run (50-100 g wet mass of cells). So is there a suggested "miniprep", or can I just do a simple cell lysis, spin down cell debris (10k G) and use the supernatant. Looking for something clean enough to put through a SPR, but equally will work in ELISA.




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