Constructing and Screening a Recombinant DNA Library 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
you could point a great professor just listening to his sound, the pitch on his voice. Like a music of science.. as if he becomes knowledge and the knowledge becomes him goes so comfortably...
This is so cool! I'm not very familiar with chemistry or biology but I'm watching all of these videos and it all makes perfect sense. He's a great instructor.
LOL I love his lecture and how he ask interesting questions that students can engage and think like a scientist by just starting with questions ! LOVE IT
You could also screen by putting each gene fragment into a vector such that it was inside a partial lacZ gene (at least in E.coli) -that is, you put the polylinker in the plasmid lacZ. You can then do a simple blue/white screening.
What does he mean by the "catalog"? Do molecular biologists look at a catalog and order DNA ligase and different enzymes that are mass produced in a lab somewhere so they don't have to make everything they need themselves? That sounds cool.
The use of the word 'catalogue' shows the age of these videos, as nowadays everybody orders their reagents online. But in essence it is the same. Yes- molecular biologists simply order enzymes they require for their cloning, such as DNA ligase, restriction enzymes and Taq DNA polymerase. In the 'olden days', labs would purify these enzymes themselves. However, given the amount of time this takes, as well as the costs involved to ensure they are of optimum quality, it is frankly much easier to just buy these enzymes in from a commercial provider.
yes, because if you are in research you can not waste time or make a mistake ....these companies dedicate to create wat you need so u just order to gain time in ur research
You cannot use that restriction enzyme, and must select another one. You can actually engineer your own restriction sites onto your insert by PCR, therefore tactically choosing an enzyme that you know will not cut inside the insert but only the regions flanking the insert.
He talked about it in a previous lecture about cloning (Basic Mechanisms of Cloning, excerpt 3) . With a restriction enzyme you put there a methylase, an enzyme which methylates the DNA here and there, and then, at random, you eventually get a cut of DNA where the gene is intact.
+Mudit Jain These clips are not complete lectures. The clips are excerpts that cover a specific concept. From the syllabus, "Fundamentals of Biology was designed specifically for independent study. It draws upon material developed for the three versions of MIT's Introductory Biology classes known as 7.012, 7.013, and 7.014." For more details and course materials (assignments with solutions, exams with solutions), see the course on MIT OpenCourseWare at ocw.mit.edu/7-01SCF11.
+MIT OpenCourseWare Thanks for the info. Are the lectures in ocw.mit.edu/7-01SCF11 complete lectures? They seem to be shorter than usual class duration of 1.5 hours. Is there any source to get complete lectures in sequence? They will be really helpful for self study.
By performing mutagenesis on yeast and then plating them out on complete media, and then replating the individual colonies on minimal media. The ones that don't grow on minimal media, are mutants. Then you can go back to your original plating to harvest the mutants.
MrRabastan One question. The intake of the plasmic DNA into the Arg-mutant yest, is it called Transfection? I'm confused with the correct term for this.