Questions tagged [transformation]
The genetic alteration of a cell resulting from the direct uptake, incorporation, and expression of exogenous genetic material from its surroundings.
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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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Griffith's Experiment Conclusions
My teacher gave us these two questions on Griffith's experiment.
How did Griffith's experiment rule out that the rough cells could have used the capsules of the smooth cells to become pathogenic?
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Terminology for Transgenes
I'm designing an in vivo delivery vector for therapeutic transgenes.
I have two different potential versions of the transgenes. If they were innate, they'd be referred to as "alleles." Does ...
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Plasmid Design and Integration events [Single vs Double cross over]
When linearising a vector by restriction digest within the middle of a homologous region can a single cross over integration event only occur if the plasmid is re-ligated within the cell after ...
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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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How much Kanamycin is too much for selection in dh5α E. coli?
I'm using Kanamycin for selection after electroporation of my plasmid containing a kanamycin-resistance gene at a concentration of 50mg/L prepared in 1000x stock that is usually kept in frozen ...
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Which method of gene amplification for toehold switches?
My team and I are from a high school and are planning to carry out some research investigating some toehold switch riboregulators which we have designed in silico. However, we have little experience ...
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How to identify the GPD gene when the sequence varies between organisms?
I'm reading a paper on genetic transformation of a fungi and the plasmid used in the paper uses two forms of the same GPD (glyceraldehyde3-phosphate dehydrogenase) promoter to drive a GFP gene, one ...
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Questions regarding transformation in bacterial cells
First off, in transformation the donor DNA aligns itself with the complementary bases in the recipient DNA.
Now a "perfect" alignment of the donor DNA ( Sorry if my terminologies are ...
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Plating 96-well bacterial transformations
I need to clone 96 different inserts into the same backbone in arrayed format. I am planning to perform Gibson assembly reactions in a 96 well plate, and then do chemical transformations in a 96 well ...
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Gene of interest number of copies in transformed K. pastoris
When transforming a K. pastoris strain, is there a way to predict how many copies of the gene will be integrated following the homologous recombination? More specifically is it something that a ...
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How does the DNA cross through bacterial cell wall during electroporation?
There exists a lot of literature on electroporation of Gram-positive and negative bacteria. Most of it gives an explanation that electroporation works by creating transient pores in cell membranes of ...
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Is a butterfly one or two animals?
I have read somewhere that a butterfly might be two animals that combined together. One animal was a worm-like creature and the other an insect.
And the insect basically hatched inside the worm. ...
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How can I improve efficiency of Ecoli transformation?
I am an intern in biology institute. I have a 17.3kbp plasmid need to transform to Ecoli. But I have tried many time but have no or very few colony on LB plate.
I know large plasmid have less ...
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are there any species that we can identify as being 'mid-way'though an evolutionary change?
I have just learned that dolphins evolved from a dog/cat like land mammal (Mesonix) that became ever-more water venturing. I understand and can visualise how the arms legs and tail slowly evolved into ...
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