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Questions tagged [competent-cells]

Competent cells are bacteria which have been treated with chemicals and which are able to take up external DNA.

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Lambda red recombination without electroporation

All protocols that I found about Lambda red recombination are using Electroporation as a method to introduce (inject) the homologous DNA (usually a PCR products or a linear dsDNA) to the E.coli cell. ...
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Cloning problem. Digestion test at the end of the cloning is always negative

I have a problem with my cloning, I'm trying to clone a 483 pb in a pSecTag vector with restriction sites HindIII and BamHI. Here are the procedures I'm doing: • I introduce the restriction sites ...
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Advantages and disadvantages of sporulation compared with competence in bacteria?

Why do bacteria have both of these mechanisms to deal with the same environmental stress: nutrient deprivation? In a culture exposed to this condition, often both competent cells and sporulated cells ...
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Why is competence useful for a starving cell?

I know that some bacteria become competent when there is not much food available in their environment. I don't really understand why this is useful for the cell. I've seen some other organisms form ...
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Problem with bacterial transformation with electroporation

I have a problem with a bacterial transformation of a yeast gene that I can not solve. I isolated yeast DNA and did a PCR to get my product. I am using pCGCUm vector with a GFP construct. I digest ...
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Is every cell competent?

I would like to know if every cell is competent or if there are any cells that are not competent.
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Why am I getting low transformation efficiency with DB3.1 E.coli cells?

I am making competent cells using DB3.1 E. coli cells. Even after following the exact protocol (Inoue method for ultracompetent cells) given in 'Sambrook and Russel', I am not getting transformation ...
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Cell growth conditions for preparing electrocompetent cells

Usually the protocol for preparing electrocompetent E. coli cells calls for growing the cells at 37deg and 225rpms until they reach OD of 0.3. I was wondering, is there any reason they should grow at ...