Questions tagged [ecoli]

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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Salvaging a plasmid from a cell culture stored at the incorrect temperature

I have E. coli with transformed plasmid on agar in a vial. It has been stored at -20 degree C without glycerol stock for 18 hours. This is a continuation of: Survival of E. coli on agar plate at -20 ...
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973 views

Survival of E. coli on agar plate at -20 degree C without glycerol stock

If I kept E. coli transformed with plasmid at -20˚C without making glycerol stock, will it survive? Actually I have kept it this way for 18 hours. What will happen with it? Will it grow in fresh media?...
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4answers
3k views

Why is it sometimes difficult to resuspend E. coli in P1?

It's a curiosity question. When I'm doing minipreps after pelleting the bacteria sometimes it's very easy to resuspend them in P1 (Qiagen kit), but sometimes they form a rubbery clump that is very ...
3
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1answer
64 views

Literature about putative epigenetic state changes in mammal sequences after cloning steps in Escherichia coli

I would like you to point me out some literature about putative epigenetic state changes in mouse/mammal sequences after cloning steps in Escherichia coli. This are the last search details I used in ...
5
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2answers
10k views

What is the appropriate method to send a strain?

We are planning to send an ecoli strain to another lab. What is the appropriate way to do so. No MTA is required but the options seem to be using filter paper or sending a glycerol stock on dry ice.
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3answers
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What makes E. coli yellowish?

LB, TB media, and yeast extract are also yellow but at the heart of the question. What chemical(s) make E. coli yellow?
5
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3answers
326 views

Measuring reverse transcriptase activity in E. coli

Goal: To quantify the HIV-1 reverse transcriptase (RT) activity for a number of mutated RTs. Idea: Put RT into a vector in E.coli. Deliver GFP mRNA to E.coli. RT will process the mRNA. The ...
9
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1answer
337 views

How long does it take for E. coli to shift feedstocks?

With our fermentations we're noticing that it takes an appreciable amount of time for E. coli (K12 variant) to change from being metabolically streamlined on amino acids to being metabolically ...
12
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3answers
6k views

How long can E. coli stocks be stored at -20°C?

I'm volunteering for a biohacker lab - biocurious in Sunnyvale. The have a pretty good set of equipment - gel boxes, incubators, but they don't have a -80°C freezer yet. I'd like to set up some ...
6
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1answer
341 views

Which cryogenic vials and caps are ideal for storing glycerol stocks?

There are internally threaded, externally threaded, natural caps. For the vials, there are skirts, star-footed, and various bottoms. Why do any of these things matter?
8
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1answer
633 views

How do you knockout an E. coli gene without disrupting the rest of the gene cluster?

I'm familiar with the method to build Keio collection using single-gene deletion with an antibiotic cassette. However, what happens when there are gene clusters or overlapping ORFs and you still only ...