Escherichia coli, a gram-negative, rod-shaped bacterium commonly found in the lower intestines of warm-blooded animals. Used extensively in laboratory research.

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Why and how is DNA synthesis so much faster then RNA synthesis in bacteria?

DNA synthesis in E. coli is 20x faster than RNA synthesis at 1000nt/s vs 50nt/s. (Mirkin'05) I find that perplexing since DNA polymerization has better proofreading than the RNA variety, which ...
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57 views

Relative densitometry from SDS PAGE

I'd like to perform densitometry on a Coomassie stained SDS PAGE gel to compare a recombinant protein's expression levels under two conditions. I'm using BioRad's Image Lab software. My questions are ...
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172 views

Can I leave BL21(DE3) cells in room temperature?

I am preparing competent cells, and I finished inoculating a single colony in SOB. It has been incubating at 37 degrees Celsius for almost 16 hours since, and it's getting very late where I live. I ...
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229 views

How to prevent e coli from clumping (for FACS)?

I'm performing FACS on e coli, but the cells are clumping together so each event is multiple cells. I ran a control where I had one flask of e coli expressing GFP, and one flask expressing RFP. Run ...
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23 views

Will bacterial promoter without either -10 or -35 sequence allow no transcription?

I have a strong constitutive promoter on a plasmid inside of E.Coli, which expresses a gene. Having either -10 or -35 sequence modified to random sequence, will it ensure zero transcription from that ...
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24 views

Recombination frequency as a function of genomic location in E coli?

Are there any databases or papers that give recombination frequency as a function of position in the E coli genome?
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96 views

Regulation of Cra protein level in E coli

Catabolite Activator/Repressor, Cra protein (formerly known as Fructure Repressor FruR) plays a significant role in central carbon metabolism of E coli. Its activity is inhibited by fructose-1,6-...
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12 views

Is there any technique other than restriction enzymes to introduce staggered cuts into DNA?

I'm curious about introducing staggered cuts into DNA, but wonder if there is a way to do this in vivo in E. coli, as I heard introducing restriction enzymes on a plasmid would end up just shredding ...
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23 views

Monod Kinetics--Growth of E.Coli in a Bioreactor

I am doing a study for a lab class. We are growing E. Coli in a bioreactor. We want to vary the volume of the initial biophase which we add after the substrate has been brought to 37 degrees C. My ...