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e coli cell lysis


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#1 zx0819

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Posted 07 July 2009 - 09:40 PM

does anyone here encounter the problem before that genes orignating from different organisms will effect the efficiency of e coli cell lysis? I am experiencing this problem right now. some of my genes from spefic species introduced into e coli but i can not break cell to get the enzyme extract by sonication, sometimes even using the lysozyme at first. What can I do to solve this problem? I hope someone here can give me some suggestions. Thanks in advance. ;)

#2 molgen

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Posted 08 July 2009 - 06:18 AM

Are you shore that you have E.coli cells?
You might have a contamination of some sort (like yeast).

#3 zx0819

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Posted 08 July 2009 - 05:02 PM

View Postmolgen, on Jul 8 2009, 10:18 PM, said:

Are you shore that you have E.coli cells?
You might have a contamination of some sort (like yeast).
Thanks for your reply, molgen.
If there are some contamination, it should not be yeast. I checked under the microscope. They look like bacteria. But I can not identify whether they are e coli or not.
If there is no contamination, what should I do?

#4 zx0819

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Posted 08 July 2009 - 07:27 PM

View Postzx0819, on Jul 9 2009, 09:02 AM, said:

View Postmolgen, on Jul 8 2009, 10:18 PM, said:

Are you shore that you have E.coli cells?
You might have a contamination of some sort (like yeast).
Thanks for your reply, molgen.
If there are some contamination, it should not be yeast. I checked under the microscope. They look like bacteria. But I can not identify whether they are e coli or not.
If there is no contamination, what should I do?
I forgot that I had a positive control and it always worked as expected.




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