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plasmid curing

plasmid curing - Molecular Biology Techniques

plasmid curing - Molecular Biology Forum. Includes forums for common molecular biology techniques.


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  #1  
Old 01-07-2008, 08:11 AM
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Default plasmid curing



Hi,I want to cure plasmids from Gram positive bacteria.Does anyone know some methods that is nontoxic and efficient?
Thank you.
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  #2  
Old 01-07-2008, 01:31 PM
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Default Re: plasmid curing

When you mean "cured" you mean removal of the plasmid?
The definition I know is: curing is the elimination
of a plasmid from its host cell.

If that is so, I found this protocol which was from an
old bacteriologist and it worked for a friend of mine
(who did it once though):
  1. Grow bacteria with plasmid overnight. use approximately 2x10^9 cells/ml.
  2. Dilute them to 1x10^4 cells/ml in LB, 0.1M Tris, pH 7.6, 20ug/ml acridine orange.
  3. Grow overnite in dark.
  4. Replica plate to find "cured" bacteria.
Also, you can use Novobiocin as it interferes with gyrase
(as does Nalidixic acid and several other agents -
Oxolinic Acid) which is needed for plasmid replication.

ethidium bromide has been used along with other intercalating
dyes.

see this paper for a new method:
[Only registered users see links. ]

It also helps to culture the bacteria at higher temperatures such
as 42C or 45C.
See this paper for more information on temperatures:
[Only registered users see links. ]

Good luck
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  #3  
Old 09-24-2008, 05:14 AM
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Default Re: plasmid curing

hi, can anyone help me about plasmid curing of gram positive bacteria using ethidium bromide?
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  #4  
Old 12-25-2008, 08:40 AM
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Default Re: plasmid curing

Hi, i want to cure antibiotic resistent plasmid from coliforms is any one know the method please help me
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  #5  
Old 09-03-2009, 06:35 PM
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Default Re: plasmid curing

Quote:
Originally Posted by qwqwe2000 View Post
Hi,I want to cure plasmids from Gram positive bacteria.Does anyone know some methods that is nontoxic and efficient?
Thank you.
kindness regard
Elham Bakhshi
Alzahra university
Tehran,Iran.
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  #6  
Old 01-06-2010, 01:30 PM
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Default Re: plasmid curing

thanks but ineed any think from curing with acredien orang
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  #7  
Old 01-06-2010, 03:26 PM
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Default Re: plasmid curing

pleas help me ineed curing by acredien orang
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  #8  
Old 01-10-2011, 09:34 AM
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Default Re: plasmid curing

Quote:
Originally Posted by curious student@biotech View Post
hi, can anyone help me about plasmid curing of gram positive bacteria using ethidium bromide?
and also for gram negative bacteria.
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  #9  
Old 01-17-2011, 12:03 PM
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Default Re: plasmid curing

thanxxxxxxxxx darling
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  #10  
Old 05-07-2011, 07:25 PM
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Default Re: plasmid curing

I am trying to get bacteria cell free of plasmid, but I could not get the protocol to do so.
Please who can help me?
Omar
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