Something that made me scratch my head: The synthesis of DNA goes from 5' to 3'. But this is on the _synthesised_ strand, not the _template_ strand. So at 1:33 he chalks in the nucleotides on the end labelled 3', but remember DNA is anti parallel and the new 5' end will line up with the old 3' end.
MrGoatflakes If you look at the figure on the board you will see two DNA strands, one in blue that is left to right running in the 5' to 3' direction and one in orange that is left to right running in the 3' to 5' direction. These are two complementary strands of the original DNA molecule that will be amplified.Each of these strands will serve as a template for the synthesis on a new strand. In each case, a primer will bind by base paring to the template strand and new nucleotides will be added to the free 3' hydroxyl of each primer. Each newly synthesized strand will be made in the 5' to 3' direction.
While PCR is exponential (a pair of different primers), Sanger Sequencing is linear and made each strand (a single primer) at a time... They are not the same, they share the fact that they are DNA synthesis based techniques...
I know I'm 8 years late to the conversation, but when you use heat to denature the DNA strand, are we assuming that helicases cannot be used? I'm just used to helicases causing the DNA strand to split rather than heat.
Kary Mullis talked out about the huge flaws in HIV leading to AIDS theory, such mistakes that’s huge percentage of the population still aren’t aware of , crazy 😜
For a second I thought he said "and their useless" at the end LOL. What he was talking about at the end sounded a lot like the Sanger method - is that the same thing or something else in this case?
Well it's not the PCR method but dideoxy termination sequencing, nonetheless it's very interesting and the teacher is so attractive that I feel quite in love with him.
What about bacterial DNA? Since it is circular, one could expect that at each PCR cycle, elongation of the new strands should stop only after the completion of the full circle. How can then we achieve isolation and replication of a particular segment? Must we first break the DNA at some point so as to transform it from circular into a linear double strand?
I know that restriction endonucleases are used to break up DNA. They have the property of snipping wherever a certain very short combination of bases appear, and which sequence they cut at depends on the particular nuclease. Many cut in such a way as to leave an overhang or "sticky end", where the double strand has a short single strand overhang. I don't know if you can use the predictable sequence at the start of the cut is long enough to make a useful primer, but if not I'm guessing what you could do is make something that complimented the sticky end, along with extra, random but known bases, amplify that once and THEN use the compliment of that as your primer for the main PCR reaction.
Hello. I don't understand one thing (so far): How do we know that this particular primer is going to "bond" with our target sequence? We need to know, on first hand, the sequence we desire to amplify. Right? So, for that, we need to know the genome. Thank you.
Você pode conhecer de antemão uma pequena região da sequência, até mesmo pelo uso de analogias em organismos modelo. Um primer costuma ter de 16 a 28 (tipicamente 20) nucleotídeos e os fragmentos que você amplifica com PCR podem ir de menos de uma centena (ex: ALU, microssatélites) até milhares de pares de base de DNA. Uma forma tradicional de revelar-se "de novo" (do zero) é clonar fragmentos via DNA recombinante. Hoje utiliza-se o Sequenciamento de DNA de Nova Geração (NGS).
Attention all professors: writing sh**out on a chalkboard and explaining it is a great way to teach! Reading some lame powerpoint for 70 minutes or so? no so great. BTW- Great explanation, clear and to the point! Thanks
I've been learning about PCR for two weeks and had NO idea what it was for.
You just summed up two weeks perfectly!
Thank you!
Pure English and simple explanation, that is why MIT is the best univ. in the world :))
This is the best explanation I've heard so far!
such a great explanation. reallly makes the whole concept clear! thanks from england
You have no idea how much you help me to understand it.
Hey guys! Try changing the speed to 1.25, and it will sound just natural without constant pauses!
Beautiful technology.
Something that made me scratch my head: The synthesis of DNA goes from 5' to 3'. But this is on the _synthesised_ strand, not the _template_ strand. So at 1:33 he chalks in the nucleotides on the end labelled 3', but remember DNA is anti parallel and the new 5' end will line up with the old 3' end.
MrGoatflakes If you look at the figure on the board you will see two DNA strands, one in blue that is left to right running in the 5' to 3' direction and one in orange that is left to right running in the 3' to 5' direction. These are two complementary strands of the original DNA molecule that will be amplified.Each of these strands will serve as a template for the synthesis on a new strand. In each case, a primer will bind by base paring to the template strand and new nucleotides will be added to the free 3' hydroxyl of each primer. Each newly synthesized strand will be made in the 5' to 3' direction.
Michelle Mischke
yes I was just momentarily confused when he said 5' to 3' and then pointed at the 3' end of the complementary strand.
+MrGoatflakes synthesized 5' to 3' relative to the new strand, not the old one.
*****
yes
+MrGoatflakes Thanks guys, that made me scratch my head aswell! :)
concise and straightforward. Easy to understand
Fantastic explanation, technical but lucid, many thanks!
That was very handy! got a Genetics test tommorrow! Thank you very much sir.
Excellent presentation. I'd always wondered how that worked. H
This is fantastic, I'm not a genetic engineer but I was able to follow your explanation.
tranks from brazil! such a great explanation!
Explanation of reading the sequence back after PCR was excellent. Did Prof. Lander deliberately miss that out !!
While PCR is exponential (a pair of different primers), Sanger Sequencing is linear and made each strand (a single primer) at a time... They are not the same, they share the fact that they are DNA synthesis based techniques...
Amazing! I finally understand the importance of ddXTP. Thanks!
Love this! It was very helpful and clear. Thank you very much!
very simple clear explanation ever . in just 8 mints . big thanks
Very well explained....well done
I know I'm 8 years late to the conversation, but when you use heat to denature the DNA strand, are we assuming that helicases cannot be used? I'm just used to helicases causing the DNA strand to split rather than heat.
Une belle vidéo en anglais expliquant la PCR. well done.
Kary Mullis talked out about the huge flaws in HIV leading to AIDS theory, such mistakes that’s huge percentage of the population still aren’t aware of , crazy 😜
HE explained it so well an 6 year old can comprehend it. GREAT WORK!!!
Awesome explanation!
Its not PCR, its Sanger method of sequencing via PCR
Still uses PCR.
watch it from The beginning
Excellent presentation!
great and simple eplanation
Thanks so much!! You are really good at explaining things!!
Wow! Thanks for such awesome explanation!
he is going 3' to 5' on the new strand being synthesised, not the template strand. He is correct
way to go man! great explanation!! nothing but respect.
Shouldn't the nucleotides be added from 5' to 3' instead of the other way around?
For a second I thought he said "and their useless" at the end LOL. What he was talking about at the end sounded a lot like the Sanger method - is that the same thing or something else in this case?
Thank you so much for this free web source! It is saving me in Biochemistry and Genetics!
LIFE. SAVER.
Really Excellent ! - Thanks very much.
excellent and very comprehensive, thank you
Thank you for a great explanation for helping me tutor kids
Amazing explanation!!!! great job!!
i rate this one 4 stars...thanks
Sanger dideoxynucleotide chain termination sequencing
you seriously saved me Man !! HATS OFF TO YOU :D
Well it's not the PCR method but dideoxy termination sequencing, nonetheless it's very interesting and the teacher is so attractive that I feel quite in love with him.
wut
this has been incredibly useful :) Robert you are my favorite!
gracias, fue de mucha ayuda!
thank you bro
I love science...
What about bacterial DNA? Since it is circular, one could expect that at each PCR cycle, elongation of the new strands should stop only after the completion of the full circle. How can then we achieve isolation and replication of a particular segment? Must we first break the DNA at some point so as to transform it from circular into a linear double strand?
I know that restriction endonucleases are used to break up DNA. They have the property of snipping wherever a certain very short combination of bases appear, and which sequence they cut at depends on the particular nuclease.
Many cut in such a way as to leave an overhang or "sticky end", where the double strand has a short single strand overhang.
I don't know if you can use the predictable sequence at the start of the cut is long enough to make a useful primer, but if not I'm guessing what you could do is make something that complimented the sticky end, along with extra, random but known bases, amplify that once and THEN use the compliment of that as your primer for the main PCR reaction.
It was a great help !!
Hello. I don't understand one thing (so far): How do we know that this particular primer is going to "bond" with our target sequence?
We need to know, on first hand, the sequence we desire to amplify. Right?
So, for that, we need to know the genome.
Thank you.
You need to know at least the start of it. Then you synthesis the compliment of that start sequence for the primer.
Você pode conhecer de antemão uma pequena região da sequência, até mesmo pelo uso de analogias em organismos modelo. Um primer costuma ter de 16 a 28 (tipicamente 20) nucleotídeos e os fragmentos que você amplifica com PCR podem ir de menos de uma centena (ex: ALU, microssatélites) até milhares de pares de base de DNA. Uma forma tradicional de revelar-se "de novo" (do zero) é clonar fragmentos via DNA recombinante. Hoje utiliza-se o Sequenciamento de DNA de Nova Geração (NGS).
great explanation! thanks
Very nice
Great Video, thanks!
THANK YOU SOOO MUUCH! easy and clear!
WOW, amazing!!!
Is the microphone in his throat?
Thanks !
AMEN! BROTHER!!!!!!! That's the freaking truth!!
thank you.. very helpful..
excellent! thank you very much.
Hello,
I have a question
By Elisa there are many positive igg with negative igm. And negative pcr? What do this mean with clinical symptoms?
Great!!!
thank you so much!
Thanks for the tutorial. :)
i understand half of the topic...from where you started about DNA structure from that point i cant understand
Wwoow great eplanation
Attention all professors: writing sh**out on a chalkboard and explaining it is a great way to teach! Reading some lame powerpoint for 70 minutes or so? no so great.
BTW- Great explanation, clear and to the point! Thanks
thanks pro
OMG THANKYOU!
Is this technique useful to test coronavirus??
Thank you! But my confusion is with the primer.. Doesn't primase sets up 5-3 prime instead of 3-5 otherwise it would be okazaki fragments?
tenx man
He is not going 3'-->5'?
So is there an Algorythm for this
cool
Nice... But why he is saying with laziness
What ward is uttered at 36 second?
"... And then four different nucleotides."
Thank you very much :-)
Thank you KnowJesusKnowPeace
An ASMR pro.
Eu queria Tanto entender ... :/ essa língua.
When it's AGCT did he say ATCT? e_____e
dis is gud
LOL, it did sound like he said their useless
In silico: th-cam.com/video/BkTRYMjyatA/w-d-xo.html
Jesus status achieved
Appreciate that wonderful explanation!!
Thanks!