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Questions tagged [plasmids]

Often circular segments of DNA that can replicate outside of the chromosomes. Generally found in bacteria.

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Plasmid Expression Vector [closed]

If I am going to transform a plasmid into a bacterium, which will then be fed to a C. elegans, the expression vector needs to be bacterial correct? The other option is to have a worm expression vector,...
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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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can mammalian expression vector be used in e coli to produce plasmid

I would like to use this plasmid (mammalian expression Flag-HA-USP53 (Plasmid #22606) - Addgene) to produce plasmid in e coli to purify plasmid. Would this work? I am starting out with this vector ...
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Can I pellet DNA back from dissolved state?

I have a dissolved plasmid pellet in water. Can I pellet it again by centrifuging it at 13000 rpm? If not, why?
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What is the nature of plasmids? [closed]

What are the plasmids composed of? Are they composed of genomic or non-genomic DNA? I guess the answer is "genomic DNA" but I'm not sure.
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In traditional plant cloning, why do we require two different vectors (plasmids)?

So I was recently taught cloning in plants and I came to wonder what is the need to first put the gene of interest in the entry vector plasmid and then the final vector plasmid before finally ...
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If the AraC protein was a gene repressor when it binds arabinose, would there be high or low transcription levels when arabinose is present? [closed]

If the AraC protein were to function as a gene repressor when it binds arabinose, would there be high or low transcription levels when arabinose is present?
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PCR and large-scale PCR

I have performed PCR with 50 μL, which showed a clear band in the gel electrophoresis. I then performed large-scale PCR with 3650 μL, which showed an unclear band in the gel electrophoresis. What can ...
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Why don't mitochondria have plasmids?

According to the endosymbiotic theory, mitochondria are descended from specialised bacteria (probably purple nonsulfur bacteria) that somehow survived endocytosis by another species of prokaryote or ...
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Full Sequence for Plasmid pMG101

I am trying to find the full plasmid sequence for pMG101. I have been looking through other papers that have sequenced this plasmid. The GenBank access numbers I got are the following: AY009372–...
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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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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 ...
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Calcium ions and bacterial transformation

What is the mechanism by which calcium ions and heat treatment allow the bacterial membrane to become permeable, allowing the uptake of plasmids?
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Why we need a plasmid for r-DNA technology [closed]

Recently I was studying Biotechnology. When I went through the texts, I had a doubt: both plasmids and gene of interest are made of DNA stretches and bacteria directly absorb plasmids in a test tube (...
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What is the significance of running an uncut plasmid on electrophoresis gel?

What is the significance of running an uncut plasmid on electrophoresis gel? In this case we are talking about inserting a gene into plasmid, which then goes under PCR and then electrophoresis.
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use of antibiotics in selection of good plasmids

I have not understood the role of antibiotic medium in filtering the bacteria containing the recombinant plasmids with the ones having normal plasmids. Since all the plasmids of a certain bacterial ...
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What placement of plasmid parts makes the most sense?

If you want to express the largest amount of a certain or a few proteins using a plasmid, does it make sense for the antibiotic resistance gene to come after genes for proteins you intend to ...
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Questions in Regards to Isolating Promotor Region for Recombinant DNA Techniques

For a while now I have been trying to isolate the promotor region of this Protein. Using sticky end PCR, I need to isolate this promotor region so it has paticular sticky ends for the purpose of ...
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Restriction enzyme digestion

A circular plasmid of 10,000 base pairs (bp) is digested with two restriction enzymes, A and B. This produces a 3000 bp and a 2000 bp bands when visualized on an agarose gel. When digested with one ...
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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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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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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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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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Cloning in a large DNA fragment into a plasmid

I need to clone in a 30kb DNA fragment into my plasmid (~7kb). Working with such a large fragment I have run into a few problems. The first was purifying it from the PCR mix used to amplify it out of ...
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Is there any commercial vector technology platform for boosting recombinant protein production?

I am a novice in biomedical industry. Our team would like to search new vector technology for boosting recombinant protein production. It's difficult to google a firm that provide those plasmids. Dose ...
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How do we regulate the production of proteins when designing plasmids?

I think it should be no surprise that I, as many others, am interested in the new COVID-19 vaccines being developed. In my region of the world there are two mayor candidates. One is mRNA based and one ...
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Induce plasmid acquisition in bacteria to make them susceptible to antibiotics

Is it possible to use plasmids as a way of creating antibiotic susceptibility in bacteria. For example; in Microbiology, an introduction 13th edition by Pearson , Page 230 "the researchers traced the ...
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Why is Homologous Recombination (HR) more frequent at long sequence repeats?

I'm studying plasmids in bacteria (E. coli), and trying to understand the well-cited phenomenon that recombination frequency increases with longer repetitive sequences. I think this also applies to ...
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Join a linear plasmid just using a primer (without ligase enzyme)

Can I use just a primer and PCR to join the cutted plasmid like this ? without using any further enzyme like ligase. After the linear dsDNA plasmid annealed into 2 ssDNA, primer will bind to middle. ...
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History of plasmid maps

To create a small history of visual representations in biology, I am seeking the earliest published papers or books that have used plasmid maps as figures ; or strategies to effectively search for ...
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Can features of modified plasmids be divided into prokaryotic features and eukaryotic features?

Here's what I understand and please correct me if I am wrong: Plasmids modified for gene therapy or genetic engineering should contain factors for certain functions in prokaryotic cells. For example, ...
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Do most bacteria have plasmids?

Do most bacteria have plasmids? I googled that and I found this Yes, Plasmids naturally exist in all bacterial cells. Here But I know some of them don't! Is this sentence scientifically correct? ...
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Can a plasmid cause cancer? [closed]

Can a plasmid cause cancer? According to wikipedia, plasmids are normally present in bacteria They (plasmids) are normally present in bacteria , and sometimes in eukaryotic organisms such as yeast [...
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