You didn't mention how to remove the parent strand, can you add one or more methods in the video describtion? Note that I'm totally proud at this video, because it is one of the best on youtube explaining site-directed mutagenesis.
i don't think it's necessary because the parent strand will be 1 between millions of modified strands after all PCR cycles. When using the plasmids for transformation, the positive clones obtained most likely will not receive the original parent strand
It is important because the parent strand could be amplified in further PCR cycles as well, there are some ways to get rid of the parental strand. A common one is the use of the restriction enzyme DpnI which can only digest methylated DNA. Methylation is a form of DNA modification which is done naturally in organisms like E.coli where most of the plasmids come from. The newly synthesized strands are not methylated.
This was by far the most clear video I have watched on site-directed mutagenesis! It simplified it down so anyone with a bit of science background can understand. Thank you! :)
I don't think I've ever watched a video by AK Lectures and not understood the concept thoroughly. I cannot say the same of other instructional videos (including Khan Academy, who I use extensively)
i seriously think...you are unique piece on the globe..bt i wonder y your videos get less views.they are worth of crosing million...may u live long and healthier
Well described! I've looked around many websites searching for a good video, and by far, you have the best explained Site-directed Mutagenesis description. Thank you so much for the time and effort. Keep up the great work you do.
Yet again, another brilliant mini lecture. Seriously, you are helping me get well on the way to completing my degree. I used your courses to help me pass my Human Biology module last semester (one of my first second year courses) and you are proving invaluable in helping me with cell biology and biochemistry. If you ever come to the UK for a holiday please let me know because I owe you a pint or 6 (especially if I pass my degree using your material to finish with a good enough degree classification to enter teacher training). Thank you so much for these, they are some of the best bio lectures I've seen on you tube (as well as Khan Academy!)
At the end of step 2, won't there be a nick in the new strand of DNA because there is no DNA ligase? Also, step 3 shouldn't occur because DNA synthesis of another strand can't occur without a reverse strand primer that provides a 3' OH.
Thanks so much for making this video.I am a Chinese student who major in biology.I cannot always understand the auditory signs with my literate faulty.
I don't understand how is the new DNA molecule circular again Does the polymerase make it circular or you're adding a thermostable ligase to bound the resulting DNA molecule and make it circular?
Sorry master but i think that the dna pol in picture 2 should attach to the 3'-OH. You draw it at 5' and it will move 5'->3' and meet the non omologus pair and cleave it with nick-translation and replace with the right base, loosing so the mutagenesis
i'll be sitting for an exam tomorrow.. A huge thank you for this video! Just wanna ask, in step 3, what will happen to 'black' single stranded DNA molecule as the DNA polymerase and dNTPs are added into the mixture? I mean, it will probably elongate the black DNA molecule as well. Just to clarify this part. Thank you :)
can someone assist me here, at exactly 8:31 he mentions that dna polymerase attaches at the 5' and starts synthesizing the template going in the 3' direction but i read somewhere that dna polymerase does not bind to the 5' site because it is INACTIVE, is this wrong?
Thank you so much for taking the time to make this video! This was the clearest way I've heard anyone describe site-directed mutagenesis
subscribed, i wish you were in my college....10 minutes of pure explanation without changing topics and adding irrelevant information. well done sir!
This video right here is a life saver. I read textbooks, papers and articles and nothing helped me understand SDM like this video.
You didn't mention how to remove the parent strand, can you add one or more methods in the video describtion?
Note that I'm totally proud at this video, because it is one of the best on youtube explaining site-directed mutagenesis.
i don't think it's necessary because the parent strand will be 1 between millions of modified strands after all PCR cycles. When using the plasmids for transformation, the positive clones obtained most likely will not receive the original parent strand
It is important because the parent strand could be amplified in further PCR cycles as well, there are some ways to get rid of the parental strand. A common one is the use of the restriction enzyme DpnI which can only digest methylated DNA. Methylation is a form of DNA modification which is done naturally in organisms like E.coli where most of the plasmids come from. The newly synthesized strands are not methylated.
This was by far the most clear video I have watched on site-directed mutagenesis! It simplified it down so anyone with a bit of science background can understand. Thank you! :)
I am a master student and This is the most well explained video I have ever seen in youtube.This video and all of the rest.Thanks
I don't think I've ever watched a video by AK Lectures and not understood the concept thoroughly. I cannot say the same of other instructional videos (including Khan Academy, who I use extensively)
Thanks Adam! Appreciate that
So true!
Fantastic explanation. You truly have a unique talent for making the complex world of biochemistry so accessible and interesting.
There couldn't be a better way to explain it. Saved a lot of time, thanks a lot!
Explained in a thorough, but simple way. Perfect. Thank you for this amazing lecture.
i seriously think...you are unique piece on the globe..bt i wonder y your videos get less views.they are worth of crosing million...may u live long and healthier
Well said
This is a portrayal of true didactic and rhetorical skill.
Well described! I've looked around many websites searching for a good video, and by far, you have the best explained Site-directed Mutagenesis description. Thank you so much for the time and effort. Keep up the great work you do.
Mohammad Suboh Thanks! Will do :)
He is a great teacher. I would never be bumped to go to a class of him.
The way you described it makes me understand even more
Really thorough and clear. Now I can plan my mutants! Thanks!
Yet again, another brilliant mini lecture. Seriously, you are helping me get well on the way to completing my degree. I used your courses to help me pass my Human Biology module last semester (one of my first second year courses) and you are proving invaluable in helping me with cell biology and biochemistry. If you ever come to the UK for a holiday please let me know because I owe you a pint or 6 (especially if I pass my degree using your material to finish with a good enough degree classification to enter teacher training).
Thank you so much for these, they are some of the best bio lectures I've seen on you tube (as well as Khan Academy!)
You were born to teach something! Thanks.
Everytime he finishes explaining something its like revealing the trick in the magic trick
He demystifies Biochemistry. Thanks a lot AK
your explanation is much better than my prof's, THANK YOU!!!
thank you so much, your clips helped me to get through my degree, i don't know how to thank you
i'm from egypt , and i want to say that you save me always before my exams .. really thanks ! you're the best teacher ever .✋
Beautifully explained by friend
After a jard struggle from almost every sourse finally this was fruitfull
You are the absolutely best!! I honestly wish you were my professor
the best videos ever honestly
Amazing! Thorough and simply explained
At the end of step 2, won't there be a nick in the new strand of DNA because there is no DNA ligase? Also, step 3 shouldn't occur because DNA synthesis of another strand can't occur without a reverse strand primer that provides a 3' OH.
I think he forgot to include the antisense primer
Best way of explaining lecture. It's help me a lot thank you for making this video
It's great. You are the best teacher.
you saved me for biochem lecture and now you're saving me for lab work i'll be doing
clear, with moderate speaking pace
U guys need captions. Love it. ❤
Thank you so much sir . You have explained the concept in very simple and interesting way .
Beautifully done! I send your videos to everyone! Thank you so much.
You are the best! :) I'm studying biotech at University level and these videos are perfect! :)
Very well explained I need to do this process for my research. Thanks for the clear explanation.
Thank you! Got the knowledge which I have returned to my lecturers
Love your lectures. it's really easy to understand!
This was an awesome video. I watched at 2.3 speed (lol) and still got everything because he is very clear
Nice explanation
Thanks you were a big help
I don't know how to thank you enough ! very helpful explanation.
Great video! You have it perfectly structured
Fantastic. You are the best teacher.
The best video on the subject. Thank you!
Answered every question I had. Thank you SO much.
Thank you Andrey!
wonderful explanation you are helping to get good marks in exames!! god bless you!!
Tank you for this amazing lecture. There is something I couldn't understand which is wouldn't the bacteria repair the mismatched base.
Thanks so much for making this video.I am a Chinese student who major in biology.I cannot always understand the auditory signs with my literate faulty.
Best Teacher Ever 🫡
Love AK Lectures! Again getting me through my degree! :-)
Thanks bro for your nice explanation
You are such a good lecturer...and a life saver too 😭 thankyou so much 🌼
I'm wondering what's the difference between #Site-Directed #Mutagenesis and #Site-Saturation Mutagenesis(I saw this on Creative BioMart)?
I don't understand how is the new DNA molecule circular again
Does the polymerase make it circular or you're adding a thermostable ligase to bound the resulting DNA molecule and make it circular?
Absolutely Phenomenal!!
You should teach professors how to teach. Thank You!
Thanks, very useful and comprehensive explanation
te amooo!!!!explicas super bien :) I hope you understand spanish! I'm watching you from Paraguay
great video, so useful.
This is the best video ever !! Thank you!
Amazing presentation, thank you. Incredibly helpful!
May I ask how to separate the sense strand from the antisense strand in the first step?
I wish you were my teacher all the time
Super helpful, thank you!
This video is GOLD!!!
great job, very helpful video.
thank you...
Thank you so much! You are a superhero
Very clear! Thank you for the explanation
Informative ❤️
This was fantastic Andrey, well done, and thank you :)
I don't get it sir only the primer we designed contain altered bases, then after primer removal old base is polymerized
Sorry master but i think that the dna pol in picture 2 should attach to the 3'-OH. You draw it at 5' and it will move 5'->3' and meet the non omologus pair and cleave it with nick-translation and replace with the right base, loosing so the mutagenesis
In Step Number 3: I can make sure to only amplify the modified strand by only adding the accroding 5'-3' Primer (TCC...)! Right?
thank you!! it explains everything in a very simple way.
thank you
Very clear!
Thanks for a great lecture! :) But just couldn't avoid noticing a misspelled word; 'mistached' instead of 'mismatched' (left bottom)!
You are amazing, sir...................
very good. perfect
so useful! thank you very much
thankq so much sir love from INDIA
thanks god for 1.5x speed but very clear explanation
Great!!! Thank you so much!!! Simple and easy!!
Mario Morales you're welcome Mario :)
Thanks so much!
i'll be sitting for an exam tomorrow.. A huge thank you for this video! Just wanna ask, in step 3, what will happen to 'black' single stranded DNA molecule as the DNA polymerase and dNTPs are added into the mixture? I mean, it will probably elongate the black DNA molecule as well. Just to clarify this part. Thank you :)
thank you so much! this will help me a lot tmw in my final exam! :)
Estephany Cortes good luck Estephany, let me know how it goes :)
beautifully explained.. the explanation is perfect... thank you so much .. :)
can someone assist me here, at exactly 8:31 he mentions that dna polymerase attaches at the 5' and starts synthesizing the template going in the 3' direction but i read somewhere that dna polymerase does not bind to the 5' site because it is INACTIVE, is this wrong?
Watched it once and got it!
Keep up that awesome work :)
In primer design, is it necessary to put the point mutation in 3' or right in the middle? Im wondering if it would have an effect at all. Thank!
Sir, what is the difference between the Site Directed Mutagenesis and Single Nucleotide Polymorphism?
Please clarify
THANK YOU A LOT
Thanks a lot.
Thank you, for this.
Thanks ,thanks
A great lecture Sir. Thank you sir
Amazing !!!
If we do this in pcr reaction chamber how can we seal up this plasmid into the circle one