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E. coli strains

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E. coli strains

E. coli strains - Molecular Cloning Forum

E. coli strains - DNA cloning forum. Discuss the cloning of plasmids, vectors, DNA, RNA, cDNA, proteins, and other molecules for expression, sub-cloning, and other methods. Also discuss digestion, ligation, transformation, plasmid preparation, cDNA library screening, and competent cells here.

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02-13-2009, 04:57 PM

niki_nje

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Join Date: Jan 2009

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E. coli strains

This is a follow up to my dead E. coli problem (posted already on the forum).

I have been using a kit from Clontech called in-fusion 2.0 for cloning, produced my clones and transformed them into their "super competent" FusionBlue E. coli cells. This worked fine and produced many colonies, but I am keen to get back to using a more common generic cloning strain used in our lab already, or common in other labs.

I have tried to transform the same clones into DH5 alpha cells, but no colonies grew. For these reactions you need "super competent" cells (according to the website), with a transformation efficiency of greater than 10^8 cfu/ug DNA (that doesn't mean anything to me, but if it does to anyone else?....).

I was wondering if anyone could suggest any strains I could try, I've been told to try Invitrogen's Top10 E. coli or Excel-1-Blue/NovaBlue (Transformation efficiency: 10^9 cfu/μg DNA).

Are there any others which could be useful?

Last edited by niki_nje; 02-13-2009 at 05:07 PM.

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« Poor Plasmid Prep Yield | Protein expression problem »

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E. coli strains

E. coli strains - Molecular Cloning Forum

E. coli strains - DNA cloning forum. Discuss the cloning of plasmids, vectors, DNA, RNA, cDNA, proteins, and other molecules for expression, sub-cloning, and other methods. Also discuss digestion, ligation, transformation, plasmid preparation, cDNA library screening, and competent cells here.