cDNA Libraries and Expression Libraries Instructor: Eric Lander View the complete course: ocw.mit.edu/7-0... License: Creative Commons BY-NC-SA More information at ocw.mit.edu/terms More courses at ocw.mit.edu
When that student said “Human cell”, i was like, nah, must be bacterial or yeast cell, then out of my expectation, the teacher went “That’s interesting, let’s put it as an option”. That is the moment i saw the light of science, education and my ignorance... Great lecture
Same. i wish my lecture actually care if student understand the material or not. all the care about if getting to the of their prepared slides that's all
Learning in the United States seems so easy. In India, in the same amount of time, you need to process five times more data due to the competition and resulting higher floor of knowledge.
@ 7:14 - 7:32 One question, I suppose we're growing Beta Globin cells for the purpose of using it? If the antibody sticks on the Beta Globin cells, these cells become un-usable? Anyone who knows the answer, pls share your knowledge.
I suppose we'd grow these cells in order to use the beta globin itself (as in the example of insulin at the end). In that case, attaching the fluorescent antibody to the cells would kill them in the process because, as he explained, you'd first have to destroy their membranes and spill out their content. Thus the cells die, but the beta globin is still usable.
thanks a lot. now I have some questions. 1) how do you destroy their cell membrane without denaturing the beta Globin protein? 2) and how do you isolate the beta Globin protein from the mess? looking forward to your reply.
Leon, Ye Lin Kaung 1) There are many ways to break down the cell's membrane (a process called "lysis"): repeated freezing and thawing, sonication, homogenization by high pressure, filtration, or permeabilization by organic solvents. The method of choice depends on how fragile the protein is and how sturdy the cells are.
Leon, Ye Lin Kaung 2) This usually depends on the mass of the protein. Anything that weighs more than 10 kDa can simply be extracted and purified by a centrifuge (either zonal or differential). Beta globin is just under 16 kDa, so that should do the thing.
Leon, Ye Lin Kaung Otherwise you have to rely on other techniques that target other characteristics of the protein: electrophoresis (separates in function of charge), chromatography (in function of mass, charge or strength of its bonds) or simply an enzyme that binds to certain proteins
I dont get this. If you wanna clone a protein from human gemone, you simply design a specific pair of primers and amplify. Why bother fluorescent ab selection?
In my understanding, flourescnt antibody will bind to the beta globin protein in the filter paper and this would tell us which cell or colony actually expressed the beta globin gene.
Right? This is MIT right? I go to UTEP (not even a tier 1 university) and I knew how to make this contruct right away and I don't even have my bachelors yet.
