Constructing and Screening a Recombinant DNA Library | MIT 7.01SC Fundamentals of Biology

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
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[@aubreyholman1951]

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.

[@JejeBabe]

im fucking jealous of those students

[@moorpmoorp]

JejeBabe I'm here because my actual professor is a PowerPoint train wreck. Dr. Lander is amazing.

[@nellik1136]

Dr. Lander, I love your lectures, learned from you a lot!!! it's and a great review for molecular biology!!

[@Wahrscheinlichkeit]

so intrigued... Thank you MIT

[@sarnaver]

I am not only learning molecular biology but also how to become a good teacher.

[@victormcarrasco4821]

This.... is astonishing!!!

[@clickbaitgod-sj9kd]

you are a legend

[@9000fail]

Have always imagined the classes to look more professional.

[@nikitaagrawal6719]

I really love your lectures

[@Indrasblade39]

Why doesn't the Phosphatase get rid of the Phosphate on both strands?

[@hamzazeraa4785]

What if mutant yeast cell that has the plasmid with arg+ gene does attack the plasmid with its own restriction enzymes ?

[@jaimehutchison175]

What happens if the gene of interest happens to contain the sequence that the restriction enzyme cuts?

[@adam4757]

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.

[@MrsZeezeee]

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.

[@alejandroportelaolcese3018]

Excelente, muchas gracias

[@zebbycat]

So helpful! Thank you.

[@jkr4807]

i don't understand a sentence : '' run it over differerent kinds of separation'' ??

[@demetronix]

and where do I get Arg- mutant yeast ?

[@gaymo69]

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.

[@Zk_Vist]

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.

[@Bob-fj7lr]

11/23/23 - 3/29

[@sashapearl8889]

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...

[@indigoblueviolet]

so true!!!!!!!!!!!!!!!!!!!!

[@monkeywithsticks]

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.

[@gillchao5458]

Thank you for this sharing! I cannot forget my last visited to MIT when I participated in iGEM. Really hope I can have the chance to study there

[@marlenesoifer7219]

This is for bacteria phages

[@ahg9843]

They are so lucky

[@crimsonghost4107]

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.

[@adam4757]

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.

[@heydyrejas1277]

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

[@TRethereal]

what is "the catalogue"?

[@prabhatkattel3166]

It's a collection of information easily found in internet.

[@ashitabisht8450]

It's a 'price list' of all enzymes and chemicals one needs in lab

[@ammarsohail7901]

Respectful Professor I love you thanks for sharing knowledge ❤️❤️❤️

[@nunitchagucci3047]

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

[@fatimaqasim7001]

After watching this lecture it becomes so easy to understand from book.

[@fatimaqasim7001]

What does catalog means there????

[@HafizahHoshni]

19 Constructing and Screening a Recombinant DNA Library Thanks !

[@Taifune81]

What does he mean by the catalog?

[@srijan8243]

I love you

[@mervdups]

It does remove both phosphates. The phosphate used in the ligation comes from the insert which has a phosphate on one strand and an OH on the other

[@DebayanRaha]

Thanks for the lecture... Love from India

[@pallab350]

The screening of ARG1 gene can be performed by using complementary Primer end with GFP gene sequence .

[@nunitchagucci3047]

He is such a GREAT PROFESSOR

[@xxkshaexx]

He really be advertising that catalog

[@rahulsurwase6627]

You are a nice teacher .....thanks

[@ScienceGeek23]

Love this level of detail!

[@manasiprasad260]

THANK YOU SO MUCH FOR THIS!

[@sxyngel]

i wish you taught at my uni

[@songthanh896]

Great lecture!

[@marlenesoifer7219]

Spelling issue

[@abdulwasay6334]

Dulcet of biology

[@muditjain7667]

Why aren't these lectures numbered properly? And are these complete lectures?

[@mitocw]

+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.

[@muditjain7667]

+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.

[@sarcasm92]

Thank you

[@erehwyrevekool]

Thanks!

[@cartior14]

The 3' end doesn't has a phosphate group has an OH group