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BL21DE3pLysS lysis

BL21DE3pLysS lysis - Protocols and Methods Forum

BL21DE3pLysS lysis - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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  #1  
Old 05-09-2006, 07:59 AM
mattia
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Default BL21DE3pLysS lysis



We work with BL21DE3pLysS and recently most of coltures lysis. We have
just change antibioticos but we have the same resultes.
Did someone have the same problem? And do you have an idea in order to
resolve it?
Thank you
Mattia

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  #2  
Old 05-09-2006, 09:53 AM
Duncan Clark
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Default BL21DE3pLysS lysis

Historians believe that in newspost
<1147161593.134484.283780@j73g2000cwa.googlegroups .com> on Tue, 9 May
2006, mattia <[Only registered users see links. ]> penned the following
literary masterpiece:

pLysS encodes T7 lysozyme so the cells are more fragile. However they
don't normally lyse whilst growing. They do tend to lyse if the induced
culture is sat on a bench for 24 hours.

If they are lysing whilst inducing then look for possibly lethal
expression. Alternatively have you changed glassware washer or detergent
and have some detergent left behind?

Do you have to use pLysS and can't use pLysE or do without the pLys
vectors altogether?

Duncan
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they go flying by.

Duncan Clark
GeneSys Ltd.
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  #3  
Old 05-10-2006, 12:20 PM
mattia
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Default BL21DE3pLysS lysis

The problem is that lysis occours before induction; we have also wash
the glassware very well but some coltures also lysis.
Mattia
Duncan Clark ha scritto:


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  #4  
Old 05-10-2006, 03:52 PM
Cornelius Fritsch
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Default BL21DE3pLysS lysis

Last time I saw that problem it turned out that bacterial stocks (or stock
solutions) suffered from a low-level phage contamination. Maxiprep
cultures lysed approximately half-way through their growth.

Cornelius

On Wed, 10 May 2006 14:20:55 +0200, mattia <[Only registered users see links. ]>
wrote:




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  #5  
Old 05-10-2006, 04:58 PM
Joe Doe
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Default BL21DE3pLysS lysis

In article <1147161593.134484.283780@j73g2000cwa.googlegroups .com>,
"mattia" <[Only registered users see links. ]> wrote:



I would suspect phage contamination. If it is phage contamination a
phage resistant strain is your best option - decontamination is
practically very difficult. Phage resistant versions of Bl21(DE3) etc.
are available from several vendors.

Roland
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