Protein expression and purification Bacteria - Origami™ B (DE3) ABCA4

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2 Discussions

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5 years ago

5 years ago by Dobrogost Maciejewski Poland

What temperature should I use for expressing thermophillic genes in E. coli?

I am currently working with E.coli and I am trying to express thermophilic genes. What temperature should I be using for incubation after IPTG induction? I have read some papers that suggested to incubate the cells at 42 degrees for a short amount of time. Is this going to affect the protein?

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Discussion

5 years ago

5 years ago by Lalli Rislakki Finland

Can IPTG in alkaline conditions become toxic to the bacteria?

Hi everyone! Currently I am attempting to measure the beta -galactosidase activity by inducing the lac operon in E.coli cells. I am trying to grow my cells in alkaline conditions with a pH > 8.5 but when I induce the cells with IPTG they are not growing. The IPTG concentration I have tried using is 0.1 and 1mM. Is there a chance that IPTG in these concentrations in alkaline conditions becomes toxic to my bacteria? Should I lower the concentration or do you think that the problem lies elsewhere?

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Found 1 matching solution for this experiment

pCEP4 Mammalian Expression Vector + Ni-NTA Agarose

pCEP4 Mammalian Expression Vector

Thermo Fisher Scientific

Ni-NTA Agarose

Qiagen

1 Relevant paper

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