Questions tagged [plasmids]
Often circular segments of DNA that can replicate outside of the chromosomes. Generally found in bacteria.
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What backbone to use for Helper dependent adenovirus (HDAd)
I have been trying to design a plasmid for a helper dependent adenovirus but while looking i wasn't able to see a backbone specifically for HDAds so I was wondering if a general adenovirus backbone ...
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Which is the best results for Plasmid dna extraction on gel electrophoresis?
I did plasmid dna extraction and uploaded on the gel.
Well 1: marker
Well 2: nicked,circular,and supercoiled bands
Well 3: linear,circular,and supercoiled bands
Well 4: circular and supercoiled bands.
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How to separate two plasmids from E coli with the same backbone?
I'm using transformation-associated recombination (TAR) in yeast to capture a biosynthetic gene cluster (32 kb) by transforming gDNA and a capture vector (11 kb) with homology arms. I identified a ...
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Problems reading my gel electrophoresis result
This is an image of my gel electrophoresis result.
I used the alkaline lysis method to extract E. coli plasmid and put it on gel electrophoresis. Based on what I had learned, I expected to see a band ...
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What are possible tweaks when a protein expression from a plasmid shipped on paper fails?
In my group we recently tried to express a protein necessary to perform an analytical method in BL21(DE3). The first two attempts did fail and no colonies did grow on the agarose. I hear, that the ...
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Can introns be removed when designing protein expressing episomal plasmids?
If your goal is to cause a specific protein to be expressed transiently via an episomally maintained gene, could you just exclude all introns and the exons not associated with that transcript variant ...
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What is the enzyme production rate of a single E. Coli bacterium?
Suppose an E.Coli bacterium intakes a plasmid which encodes for an enzyme, if the promoter strength is known, is is possible to predict the rate at which the enzyme is produced? Is there an equation ...
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Migration of cut vs. non-cut plasmid on gel
As you might know there are 3 common forms of a plasmid:
ccc-form (CCC), being supercoiled.
oc-form (OC), being nicked and therefore relaxed.
linearized form (L), being cut on both strands and linear....
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What is a good expression vector for Salmonella enterica subsp. enterica serovar typhimurium
I am interested in expressing custom proteins in a Salmonella strain, however I am facing difficulties in finding the appropriate expression vector for it. It seems that most of the resources provide ...
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Linearising plasmids for CRISPR experiment
I am currently designing a mock CRISPR knock-out experiment, and I’m wanting to insert a plasmid for selection. Using a restriction enzyme at 2750bp for cutting, would the location of the cut site ...
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Production method of custom seed mir451? How to edit middle of a fragment?
I am trying to create a plasmid that produces mir451 targeting a new gene, so I need to alter the seed sequence. The seed sequence is 7 bp long, so I think that is too long for PCR mutagenesis. How do ...
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Can the genetic sequences for CRISPR components be inserted into the host genome so that the cell perpetually produces CRISPR components?
Theoretically speaking, can you insert the gene sequences for cas9, sgRNA, and promoters into the host genome so that the cell perpetually produces CRISPR components?
In this scenario, I'm guessing ...
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Cut plasmids with unknown sequence only once
Is there a way to linearize plasmids in a sequence-independent manner?
The circular plasmid should be cut ideally only once. It does not matter where. But as soon as it is linearized, it should not be ...
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Plasmids for high school students?
For a final project in AP Biology, I am planning to design an experiment that uses the plasmid #170380 from Addgene. However, under availability they say "Academic Institutions and Nonprofits ...
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Why does plasmid recombination require precise "copying and pasting" but microinjection doesn't?
Just learning about biotechnological techniques of gene transfer at the moment. With bacterial plasmid recombination, from a high-school level, we are taught that:
The desired gene is cut using a ...
