Gene is gone

About microscopic forms of life, including Bacteria, Archea, protozoans, algae and fungi. Topics relating to viruses, viroids and prions also belong here.

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jacksonchen
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Gene is gone

Post by jacksonchen » Fri Jul 30, 2010 9:27 am

Hi,
Here is the problem. I got a gene fragment from a bacterium by using specific primers for PCR. The band was clear and single in agrose gel. But after cultivating this bacterial strain for several generations, I could not detect this gene any more. I had tried to check every step. The primers were fine. The genomic DNA is also good enough. Even there was no problem with PCR machine. But I still got nothing from PCR. Then I guessed that the strain might be mutanted. So i cultivated the original stain which was preserved in glycerol at -30℃. But still no results. This is driving me crazy! Hope to get some suggestions from you guys. it would be appreciated!

Jackson

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JackBean
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Post by JackBean » Fri Jul 30, 2010 10:54 am

Wasn't the gene on plasmid?

If you get no signal even from the original strain, then must be problem somewhere after isolation...
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

jacksonchen
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Post by jacksonchen » Mon Aug 02, 2010 9:20 am

Thanks, JackBean. There was no plasmid in bacterial genome. Is it OK to preserve the strain in in glycerol at -30℃?

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JackBean
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Post by JackBean » Mon Aug 02, 2010 11:46 am

yeah, we usually keep them at -80°C, but I guess that should be fine too. What was that for gene? Can you detect the enzyme activity?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

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