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If you have 2 different mutations (irrespectively of where they sit in regards to the final gene-product), and both correlate perfectly with a given phenotype, (and you have dismissed any other potential cause), you can not formally say, if the first mutation, or the second, or both mutations are required for your phenotype. (Though you can make educated ...


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Run it out on a gel after digestion and extract the vector only. Treat with a phosphatase after digestion. This will prevent religation since you need at least one of your fragments to be phosphorylated for the ligation to work. (A must regardless) sequence several colonies at the end. I recommend doing all three. Edit: option 4: digest with a third ...


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It requires two genetic constructs to create a conditional KO organism. One parent with a transgene that expresses a DNA recombinase (such as Cre) after the promoter of a gene that is ONLY expressed in your tissue of interest. The other parent contains a modified gene of interest to be removed in the presence of said DNA recombinase but is fully functional ...


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You are correct that brachydactyly is a dominantly inherited disorder and is usually caused by mutations in the BMPR1B gene. However, not everyone who has the mutation, will have clubbed thumbs (the phenotype). This is due to a phenomenon known as penetrance and expressivity. Penetrance essentially is an all-or-none phenomenon whereby certain individuals ...


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Gene editing is not done on a single cell but a bunch of cells. However, it still does not affect all the cells. Gene editing/therapy for the entire organism works in these cases: Stem cells are edited, which give rise to many other somatic cells. Usually done in case of gene therapy in which the stem cells are obtained from the patient, edited and re-...


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A general note on phenotype I think that the meaning of phenotype has become more complicated over time compared to what it was when it was first coined. As you said, in simple words, a phenotype is an observable characteristic of an organism. However, for Mendel, an observable characteristic would have been only an obviously gross change in the morphology ...


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Yep, that is correct in general. However, sometimes proteins indirectly determine phenotype. For example, the skin colour is dependent on melanin (not a protein) but the melanin producing cells work because of proteins. Further, melanin production is also controlled by proteins. Thus, skin colour is not DIRECTLY determined by proteins but because proteins ...


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There are protein complexes which are made up of multiple subunits. Whether you call that one protein or a multi-protein complex might depend on whether you have a gene-centric or protein-centric point of view.


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The plasmid won't religate efficiently if the ends are not compatible. However, it's possible that very rarely your plasmids may re-ligate, perhaps because of a tiny level of contamination with exonuclease that trims back the ends to be blunt. Or there may be spontaneous blunt breaks or other strange events. This may happen in a tiny fraction of the ...



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