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lactobacillus species plasmidIsolation ? - help me !


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#1 anonymous

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Posted 20 May 2001 - 09:00 PM

Hi !!! everybody....

I need plasmid isolation method of lactobacillus species(LB).

Lactobacillus isolation is hard.

problem wasn't destroy of cell wall(step I)

everybody help me!!!

If you have essencial destroy method, send of me

And full method .....send of me

waiting !!!!!



#2 David

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Posted 25 May 2001 - 09:00 PM

OK here is help. There are a number of steps you can use but two of these are the most relevant.

1. Add glycine to the growing culture when the A600 is = 0.9 and at a final concentration of 50mM. When you pellet the culture after 4 hours, this cell pellet can be totally lysed in suspension by lysozyme(0.1 mg/ml), proteinase K (300ug/ml) and 0.1 %SDS.

2. Use French Press ( 15000 Psi ) for lysis.

Good luck

Davidhttp://www.dstf.bigstep.com===========Playing career chess with bright minds.......every day---------------------------------------------------


#3 tomgar

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Posted 18 December 2004 - 08:06 AM

Try this: http://www.pubmedcen...=1&artid=182348
GOOD LUCK
Sorry for late answer




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