I've watched 3 other videos on this, plus my professor's lecture, and read the textbook... I was on the verge of crying because I felt so dumb for not understanding it and only your video made something click in my brain!!! Thank you so much really, you have a gift for teaching science
Good sir, I just want to let you know that your explicit lecture didn't just save me, but also many of my fellow classmates who rely largely on my detailed notes to understand this concept. I cannot express enough of how appreciated I am.
The most amazing lecturer EVER !!!! I cant thank you enough, i bet i will have watched every single one of ur videos by the end of my first biomed year ;)
Even though 2 years have gone by since you published this I feel compelled to say THANK YOU! I stumbled on your video after looking at another 6 much shorter videos which left more questions than answers. While it firstly seems you repeat concepts it is actually VERY useful to follow all of the reasoning. My Genetics was studied back in the 80's (I am a medical doctor) and your video is just what is needed for a good intelligent refresher to our current knowledge.
I'm a Premed student and can't understand genetics anywhere more than from your channel. You are amazing! I wish you make more videos on higher-level, enhanced and new genetic concepts for students of master of genetic counseling and geneticists.
Genuinely thank you SO much for ur videos, don't know how manage to make these complicated concepts look soooo easy but u just do. I remember them for ages after studying them from u
Hi, Andrey! I am doing Molecular Pathology subject at uni now. It is very complicated..and your short lectures help a lot! Thank you so much for that! Big hug!!! :)
how many cant wait to finish school, get a job, come back and than this guy. now in my fourth year Bsc Biotechnology and AK Lectures team have been making my life easier
Dude.. Your lecture just gave me that chill of discovering how awesome science is.. It hasn't happened to me for a long time ever since I took this damn MCB course.. Thank you so much..
Thanx to AK class for providing such clear concepts... I was finding for a splendid bio channel on youtube and i hope i've got u... Thank u sooo much... :-)
Thanks for your awesome job. I have a question. Can we label only primer or any other substrates (i.e dNTP , ddNTP)? I am asking this question because some sources say - It is the ddNTP that is to be labeled , some other say it is dNTP. Reviewing all of them, I have come up with conclusion that, we can label any substrate (dNTP, ddNTP or primer). Am I right?
Hi , the lecture was superb. I have one doubt. When we dont know the dsDNA seq, then how we can design the primer complementary to the 3' prime end? looking forward for your quick reply
Grat video, well explained, thanks. My question is how can I have the primer complementary if I have to find the DNA sequence? Do we use database or other? If we use database, how did the biologists when they didn't have any information? Sorry for my english :)
+random we would cut the DNA with a restriction endo nuclease enzyme and which has a property to cut DNA at specific sites.... like BAMH1 , HIND1, etc....
We can just add a segment of DNA that we know it sequence by clonnage method while we may produce a primers are complementary to that sequence . So we can sequencing the unknown DNA sequence. Sorry for my English 😊
How is the intended DNA sequence isolated in the first place? Does this method require amplification of the DNA before addition of the nucleoside triphosphates?
How can you determine the sequence in the beginning of the strand to make the primer? 10:41 you said there is a possibility that it doesnt use the ddatp in strand 2, and instead is used in strand 3 Also, if only strand 1 and 3 were to be produced, how would you determine the skipped A in strand 2?
Andrey, is there a possibility that we could also get an entire chain of the DNA molecule also (one that does not produce any fragments) because the 1% ddNTP may not cleave any nucleotides?
I dont understant well how from 1 chain of DNA I can finally get 3....so tha's mean that primer need to settle down more times than onces?? I dont understand how from 1 chain I can get more fragments.
how do we typically find out what the sequence is of the end of the DNA strand? You mentioned that this is something you must know beforehand in order to use this technique. However, how would we find out what the sequence of the end is given that we don't know what it is?
Thank you for your explanations, I was wondering would there be many fragments of the same template DNA in the mixture in order for Step 3 to occur? If so, which step does the amplification of fragments occur, is it PCR before the sequencing takes place? Thank you!
I have a question in regards to Sanger Sequencing. Sequencing of the DNA fragments will stop at all possible positions where the particular dideoxynucleotide is incorporated before starting a new chain and proceeding to the next ddNTP. How is this ensured exactly? How do we not end up with whole lot of random sequences or all short fragments or all really long fragments of DNA for each ddA, ddT, ddC and ddG? I hope my question is clear, Thank you.
student's opinion, i think its achieved through the design of the whole procedure: on the ddNTP is the 3' position OH lacking an oxygen atom, therefore a phosphodiester bond cannot be created between the 5' phoshate of the next dNTP and the incorporated ddNTP
I love the videos, brother, but I have a quick question. Shouldn't we get one more band in each SDS channel corresponding to the unlikely scenario where the entire strand is synthesized via dNTPs?
Theoretically, if an entire DNA strand were synthesized using only regular deoxynucleotides (dNTPs), without encountering any ddNTPs, you would expect a band representing the full-length sequence in each lane (corresponding to A, C, G, T) of the gel electrophoresis. However, it's exceedingly unlikely that the polymerase would continuously incorporate only dNTPs without encountering and incorporating any ddNTPs during the sequencing reaction. The random integration of ddNTPs ensures that termination occurs at various positions throughout the sequence, creating terminated fragments of different lengths. This randomness makes it highly improbable to obtain a band representing the full-length sequence in a single lane of the gel, as the likelihood of the polymerase completing the entire sequence without encountering a chain-terminating ddNTP is extremely low.
how many single-stranded DNA molecules are we putting into those beakers? and.. what if we put too much DNA strand molecules and the ddNTP and dNTP have been used up ( causing repeated base-pairings of the same mass) before the template strand can be sequenced completely?
Sir many many thnak u which i can't explain here.❤❤ This video helped me a lot..after Professor & many video lecture, finally i understood Sanger method by ur lecture....... 🥰🥰🥰 Tnq tnq tnq tnq tnq tnq🥰🥰🥰🥰🥰
What is the difference between fluorescent dye tagging and radioactive labeling, other than the fact that one uses UV light and the other uses autoradiography?
Very helpful.. thanks! But what if the DNA pol. didn't incorporate any ddNTP? In this case we will have 4 fragments...right!? pls. respond I have exam tmw.
I've watched 3 other videos on this, plus my professor's lecture, and read the textbook... I was on the verge of crying because I felt so dumb for not understanding it and only your video made something click in my brain!!! Thank you so much really, you have a gift for teaching science
Of the 3 videos I've watched on this, only AK managed to clearly explain the concept. Thumbs up.
When I am done with medical school, you're getting a good donation!
So, are you done yet?
ME TOO
AK lectures are absolute life-savers!!
@Alyssa Colbert good luck!
My thoughts exactly. Same goes for Nancy Pi
I always come back to this video before my Genetics tests and exams - it's a faster way to learn it then reading through piles of notes. Thanks!
Good sir, I just want to let you know that your explicit lecture didn't just save me, but also many of my fellow classmates who rely largely on my detailed notes to understand this concept. I cannot express enough of how appreciated I am.
The most amazing lecturer EVER !!!! I cant thank you enough, i bet i will have watched every single one of ur videos by the end of my first biomed year ;)
I have been following you, and all I can say is you are a genius. You teach with clarity and simplicity
YOU!!!!! You are the one to finally get this into my head. Thank you so much for this video, you're a legend.
Even though 2 years have gone by since you published this I feel compelled to say THANK YOU! I stumbled on your video after looking at another 6 much shorter videos which left more questions than answers. While it firstly seems you repeat concepts it is actually VERY useful to follow all of the reasoning. My Genetics was studied back in the 80's (I am a medical doctor) and your video is just what is needed for a good intelligent refresher to our current knowledge.
Professional, Clear, Concise, and Thorough. Thanks for the great video!
I've watched many Sanger sequencing videos. This one is my final stop. Thanks Sir!
these are by far the best videos for bio/ med students on youtube - we can't thank you enough!!!!
This was incredibly well-laid out and helpful! Thank you for going through a complete example from start to finish.
You are honestly the most amazing teacher. Thank you, since it's rare that I can follow perfectly.
This is the most understandable video I have ever seen. Thank you, Andrey!!!
Thank god for your simplicity! You just made my presentation for Sanger vs next gen extremely easier! Thank youuu! Keep it up!
This is the only video on you tube which really explain the sequencing. AK you are too good.
That was so clear. The flow in your explanations worked so well i could easily follow every word you saying. Thanks a lot.
The type of a teacher I wish I had for the rest of my studies... thank you very much!
God Bless you man. It's hard not to learn something new watching your lecture. Your channel will be my winter break!
Honestly you always save my life i can't tell you how much i'm thankful for all your lectures
I'm a Premed student and can't understand genetics anywhere more than from your channel. You are amazing! I wish you make more videos on higher-level, enhanced and new genetic concepts for students of master of genetic counseling and geneticists.
One like is not enough for this video. This is hands down the best video on this topic!!
Can't explain how helpful these videos have been! Thank you, thank you so much!!
AK lectures are fantastic.Each time I'm in problem you are the one who solve it.
This is the best one out of 4 videos I watched. I was so lost. Thank you so much G.
Genuinely thank you SO much for ur videos, don't know how manage to make these complicated concepts look soooo easy but u just do. I remember them for ages after studying them from u
I'm binging on your channel in preparation for my bio-chem final tomorrow morning. Thanks!
so easy, when explicated by dude like you. peace from Morocco
Hi, Andrey! I am doing Molecular Pathology subject at uni now. It is very complicated..and your short lectures help a lot! Thank you so much for that! Big hug!!! :)
Thanks Sveta! Good luck with the course! study hard :)
I just signed in to like this video because this video deserves to be liked and commented.
how many cant wait to finish school, get a job, come back and than this guy. now in my fourth year Bsc Biotechnology and AK Lectures team have been making my life easier
OH MY !! THANK YOU SO MUCH !! I FINALLY GET IT, it's so clear
Sir, please upload some videos on the Next Generation sequencing , that would really be a great help... Your lectures are as always AWESOME.
wow i thought this was way more complicated than it looked. Thanks for making it simple
Good work! Watched the channel during undergrad, then masters and here I am while doing PhD!
Thank you for being a long-time viewer! Best of luck with your PhD!
The best explanation on youtube. Thanks!!!!
+Dill Mart Thanks!
Thanks 🙏
it can't be better thanks a million I was kinda lost, your lecturer makes it clear to me
Just perfect as always, AK!
You saved my life. I have been struggling with this
Dude.. Your lecture just gave me that chill of discovering how awesome science is.. It hasn't happened to me for a long time ever since I took this damn MCB course.. Thank you so much..
Also I played this video at 2.0 speed and max volume, which kept me concentrated af lol
Oh thats pretty amazing Tommy; that chill is hard to come by, glad this content gave you that! Keep at it and best of luck in your MCB course!
+AK LECTURES (Andrey K) Thank you!
Thanx to AK class for providing such clear concepts... I was finding for a splendid bio channel on youtube and i hope i've got u... Thank u sooo much... :-)
Great video! Very clean job.
Thank you!
Greetings from Italy!
thank you so much, so comprehensive. I wish you the best on your study and career
I love this man. God bless you!
Wonderful,I had doubt about this from the beginning!😢you made it so clear👍👌👌👌😊☺️
You are a great help for my BSc in Sport Sports Science.
Love you man.. You're a gentleman and a scholar.
i am such a fan of yours ... your way of teaching is beyond the top... great job thank you
Thanks Anna! glad you enjoy it!
Thanks for your awesome job. I have a question. Can we label only primer or any other substrates (i.e dNTP , ddNTP)? I am asking this question because some sources say - It is the ddNTP that is to be labeled , some other say it is dNTP. Reviewing all of them, I have come up with conclusion that, we can label any substrate (dNTP, ddNTP or primer). Am I right?
Thank you for enlightening me :)
thanks to your video tutorial I'm going to pass my molecular bioinformatics exam :)
This video was awesome! You saved my grade.
You are great at teaching! Please never stop making such videos! Awesome style! :)
lecture kha naday.
I love your videos so much. I understood my subjects very well during biomed 1st year
You make it so simple! Thank you!!
You make me pass the exam..Thank you..
Hi , the lecture was superb. I have one doubt.
When we dont know the dsDNA seq, then how we can design the primer complementary to the 3' prime end?
looking forward for your quick reply
The best video about this topic
That made sense. Thank you!
you are very good at explaining . Thank you so much , this really helped ALOT
thanks.... it was really helpful.... I had cleared my issues about this topic with this vedio.... thank you for the good work
you are the best thing that has happened to biology students ok
Grat video, well explained, thanks.
My question is how can I have the primer complementary if I have to find the DNA sequence? Do we use database or other?
If we use database, how did the biologists when they didn't have any information?
Sorry for my english :)
I wonder this too, how do i we choose a complementary primer when the whole purpose of this is to find the DNA sequence which we do not know.
+random we would cut the DNA with a restriction endo nuclease enzyme and which has a property to cut DNA at specific sites.... like BAMH1 , HIND1, etc....
May I ask about the database are ???
We can just add a segment of DNA that we know it sequence by clonnage method while we may produce a primers are complementary to that sequence . So we can sequencing the unknown DNA sequence. Sorry for my English 😊
I am so grateful for this video.
Thank you for clearly explaining this process!
How is the intended DNA sequence isolated in the first place? Does this method require amplification of the DNA before addition of the nucleoside triphosphates?
How can you determine the sequence in the beginning of the strand to make the primer?
10:41 you said there is a possibility that it doesnt use the ddatp in strand 2, and instead is used in strand 3
Also, if only strand 1 and 3 were to be produced, how would you determine the skipped A in strand 2?
Perfect, clear and concise. Thanks!
Thank you,this was an amazing! helped me understand this much better
I can't thank you enough for such great videos
Andrey, is there a possibility that we could also get an entire chain of the DNA molecule also (one that does not produce any fragments) because the 1% ddNTP may not cleave any nucleotides?
The best explanation of sanger sequencing!
I dont understant well how from 1 chain of DNA I can finally get 3....so tha's mean that primer need to settle down more times than onces?? I dont understand how from 1 chain I can get more fragments.
You are a life saver! thank you so much
Fantastic explanation, very clear and easy to follow.
how do we typically find out what the sequence is of the end of the DNA strand? You mentioned that this is something you must know beforehand in order to use this technique. However, how would we find out what the sequence of the end is given that we don't know what it is?
Yeah, I'm wondering this as well? How do we know what the primer sequence should be?
we cut the dna from a specific endonuclease so we know where it cuts the dna and therefore know end sequence
Your videos are so helpful! Thank you
Thank you for your explanations, I was wondering would there be many fragments of the same template DNA in the mixture in order for Step 3 to occur? If so, which step does the amplification of fragments occur, is it PCR before the sequencing takes place? Thank you!
Really amazing , I can get the information and concept clarification easy wow really thumps upppppppp
AMAZING explanation.
I like the way you teach, great job brother!
I have a question in regards to Sanger Sequencing. Sequencing of the DNA
fragments will stop at all possible positions where the particular
dideoxynucleotide is incorporated before starting a new chain and
proceeding to the next ddNTP. How is this ensured exactly? How do we not
end up with whole lot of random sequences or all short fragments or all
really long fragments of DNA for each ddA, ddT, ddC and ddG?
I hope my question is clear,
Thank you.
student's opinion, i think its achieved through the design of the whole procedure: on the ddNTP is the 3' position OH lacking an oxygen atom, therefore a phosphodiester bond cannot be created between the 5' phoshate of the next dNTP and the incorporated ddNTP
how do you determine what sequence is on the original strand to build the complementary RNA primer sequence?
Exactly what I want the know
@@sputnikaguya the same also... i dunno how did we make primer comolementary to the DNA template and we don't know the DNA sequence !!??
Great question! I'd like to see a video on that as well
Yep bro i also got the doubt in that
Few known nucleotides are added to the original strand so as to make the primer.ok
Wowww! You are such a excellent teacher!
thank you for benefit VDO clip. Tomorrow i am going to do the final test in microbial genetics.
So how would you know what the 3' end of the strand is to create the primer?
I love the videos, brother, but I have a quick question. Shouldn't we get one more band in each SDS channel corresponding to the unlikely scenario where the entire strand is synthesized via dNTPs?
Theoretically, if an entire DNA strand were synthesized using only regular deoxynucleotides (dNTPs), without encountering any ddNTPs, you would expect a band representing the full-length sequence in each lane (corresponding to A, C, G, T) of the gel electrophoresis.
However, it's exceedingly unlikely that the polymerase would continuously incorporate only dNTPs without encountering and incorporating any ddNTPs during the sequencing reaction. The random integration of ddNTPs ensures that termination occurs at various positions throughout the sequence, creating terminated fragments of different lengths.
This randomness makes it highly improbable to obtain a band representing the full-length sequence in a single lane of the gel, as the likelihood of the polymerase completing the entire sequence without encountering a chain-terminating ddNTP is extremely low.
Another perfect video. Thanks!
Hellishly helpful ! thank you so much
right now you're my superman :)
Thanks a lot !
Huge respect from india!
how many single-stranded DNA molecules are we putting into those beakers? and.. what if we put too much DNA strand molecules and the ddNTP and dNTP have been used up ( causing repeated base-pairings of the same mass) before the template strand can be sequenced completely?
Excellent. Love your vids man.
thank you man. saved my day!
Shouldn't there be another fragment formed for when the dNTP is used throughout the strand?
We are doing this to sequence the unknown DNA fragments, then how will we know the primer sequence to prepare them??
Sir many many thnak u which i can't explain here.❤❤
This video helped me a lot..after Professor & many video lecture, finally i understood Sanger method by ur lecture....... 🥰🥰🥰
Tnq tnq tnq tnq tnq tnq🥰🥰🥰🥰🥰
Superb video. Thank you so much for creating this content.
What is the difference between fluorescent dye tagging and radioactive labeling, other than the fact that one uses UV light and the other uses autoradiography?
Very helpful.. thanks! But what if the DNA pol. didn't incorporate any ddNTP? In this case we will have 4 fragments...right!? pls. respond I have exam tmw.