Next Generation Sequencing (Illumina) - An Introduction

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  • Опубліковано 16 гру 2024

КОМЕНТАРІ • 227

  • @ammaralramdhan8214
    @ammaralramdhan8214 3 роки тому +390

    Who else came here cause the illumina video was confusing

  • @bobu5213
    @bobu5213 Рік тому +10

    Finally somebody explaining how the technique actually works. Over videos are so vague and assume you already understand the function of all the parts. Thanks so much

  • @ericchen8287
    @ericchen8287 2 роки тому +122

    One thing to keep in mind is that adaptors are stuck with the 5’ end to the flow cell so the polymerase can synthesize DNA in 5’ to 3’ direction

    • @foodlover8708
      @foodlover8708 Рік тому

      Ooooh right thank u

    • @diego80mi
      @diego80mi Місяць тому

      shouldn't it be the other way around? DNA polymerase synthesize from 5', so the adaptor stuck onto the plate should be at the 3' end, shouldn't it?

  • @sonya1500
    @sonya1500 3 роки тому +46

    all the other vids on youtube just kinda goes on a rampage abt how innovative and amazing it is like a promotion or smth. So thank u so much my guy

  • @almudenaotalora8259
    @almudenaotalora8259 3 роки тому +301

    Incredible well explained. Non of my uni professors have been able to explained that well

  • @suranjanasikdar6224
    @suranjanasikdar6224 2 роки тому +27

    This is the best explanation on NGS. I had no clue, done so many researches but cant understand clearly. But this explanation is just awesome. Thank you so much.

  • @bertramaudiel5052
    @bertramaudiel5052 6 місяців тому +4

    A perfect and clear explanation for the NGS!

  • @Goldbio
    @Goldbio 3 роки тому +22

    Just want to give you a shout- out and thank you for such an easy-to-understand overview of this process. It is definitely helpful.

  • @kairenyeong7915
    @kairenyeong7915 3 роки тому +19

    I wish I found this video earlier lol, spent 2 semesters on this and none of my professors explained it well enough
    Now everything make sense to me

    • @DelasVC
      @DelasVC 3 роки тому

      yeah, unbelievable how much that matters and how few people are actually competent to teach..

  • @jaysung3354
    @jaysung3354 2 роки тому +8

    thank you very much sir. This is more well explained than the official Illumina videos.

  • @hashrafi7148
    @hashrafi7148 3 роки тому +22

    simple, clear, and comprehensive! Great job!

  • @noorkhan6486
    @noorkhan6486 3 роки тому +20

    Excellent, very well explained. Clearing all the concepts one by one

  • @litchfirmian7562
    @litchfirmian7562 2 роки тому +5

    Thank you, your animation and explanation are very clear. I finally understand what happened.

  • @VipulSinghal-k9c
    @VipulSinghal-k9c Місяць тому +1

    This is truly incredible. Good work.

  • @Qui-Gon-Jinn
    @Qui-Gon-Jinn 2 роки тому +3

    Mein Professor hat dein Video für die Erklärung genutzt. Sehr stark, weiter so!

    • @henrikslab
      @henrikslab  2 роки тому

      Darf ich fragen an welcher Uni, welcher Professor? :D

  • @hananghozlan
    @hananghozlan 3 роки тому +9

    this is incredibly clear and easy, thank you very much

  • @BenWinney
    @BenWinney 4 роки тому +4

    Great video. The only one that allowed me to understand this difficult topic

  • @RahulMPrathap
    @RahulMPrathap Рік тому +1

    This is the best and simple explanation ever in this topic. Great job.

  • @henrikslab
    @henrikslab  Рік тому +9

    From now on you can become a member of this channel! You will have access to some cool emojis like or .
    This is of course voluntary, but the financial support helps me a lot.

  • @nevanoconnell3356
    @nevanoconnell3356 8 місяців тому +2

    I have heard some conflicting ideas on what Hybridisation is. One thing I read seemed to imply it was the process of binding the oligonucleotide adaptors to the cut up DNA, however is that not just library formation. Then other places and your video have implied it is when the oligonucleotide adaptors bind to the complementary oligonucleotides on the flow cell.

  • @haminh6382
    @haminh6382 3 роки тому +2

    I have a question, so at the end of the vid is DNA read of only 1 small cut fragment, how do we combine all fragments that have been cut at 0:31 1 little fragment is analyzed by overlaid fragments result from PCR, then what about the whole thing, I mean restriction enzyme will cut it nicely and no overlaid could happen

  • @galafarah3377
    @galafarah3377 27 днів тому +1

    God bless you, Henrik!

  • @mrinalshrestha7442
    @mrinalshrestha7442 9 місяців тому +1

    Great video! Crisp explanation! Much appreciated!

  • @chuihoi1423
    @chuihoi1423 2 роки тому +4

    Thank you. Simple and clear. Now I know how it works!

  • @madscientist9288
    @madscientist9288 7 днів тому

    Great explanation - simple and effective!

  • @elias1376
    @elias1376 3 роки тому +3

    this should be illuminas explanation video. very well done 👍🏽

  • @ChrisFariaGTARealtor
    @ChrisFariaGTARealtor Рік тому +1

    Very cool and informative Henrik 👏👍🙏

  • @k14d57
    @k14d57 3 роки тому +4

    Besser als mein Prof. es je erklärt hat, Danke für das Video :)

  • @amanaparbin4390
    @amanaparbin4390 3 роки тому +4

    Such a amazing esiyer undesirable way of teaching ❤️ we need teacher like u . Great 👍🙏

  • @卢彦宏-k5o
    @卢彦宏-k5o 3 роки тому +4

    clear and easy to understand. Thank you !

  • @jonnathan.mtyyyy
    @jonnathan.mtyyyy 2 роки тому +1

    sehr gut gemacht hinnerk

  • @Korean_Dawin
    @Korean_Dawin 6 місяців тому +1

    Your lectures are wonderful!!!

  • @christianchukwujindu5133
    @christianchukwujindu5133 2 роки тому +3

    This is indeed good and helpful, thank you so much

  • @animasally4966
    @animasally4966 2 роки тому +2

    Remarkably explained! Tho I have a question. If we want to seq the foreign DNA in one go...why do we discard the Reverse based strands, cos if we would keep them, the sequencing will only mark out the initial forward strand and that's what we are aiming for! Still confused.

  • @zunairaa.9241
    @zunairaa.9241 2 роки тому +3

    your explanation was perfect but i'm definitely failing my exam tomorrow

    • @henrikslab
      @henrikslab  2 роки тому +1

      If you still read it: All the best for the exam!

  • @kidraul01
    @kidraul01 3 роки тому +3

    Incredibly clear ! Thank you !

  • @Kat-lx7qq
    @Kat-lx7qq 2 роки тому +1

    thank you so much Henrik. This was perfect!

  • @hafsahimaan2872
    @hafsahimaan2872 4 роки тому +7

    Thank you so much, you explained it super clearly!

  • @95jazmyn
    @95jazmyn 2 роки тому +2

    This was an incredible explanation! Thank you so much

  • @sonikj5568
    @sonikj5568 3 дні тому

    Thank you!! This was so helpful, my uni lecture was quite confusing and the illumina video just made it so much worse lol

  • @jessylemoine7694
    @jessylemoine7694 2 роки тому +1

    You save my exam thank you ❤️❤️

  • @tobiasr.205
    @tobiasr.205 3 роки тому +3

    Very good explanation, but I have one little question: How can it be that the two different Adaptors bind to the two different ends, and not one adaptor to both ends?
    Is the cutting done by a restriction Enzyme, and therefor it always has the same Nucleotides at the end?

    • @animasally4966
      @animasally4966 2 роки тому +3

      1. It's cos one is Forward primer and the other is reverse primer that's why they join different ends.
      2. Yes the restriction is done by a single restriction enzyme, so that the sticky ends are poduced uniformly, and yes the overlapping sequences indicate that they are cut by Restriction endonuclease that's why they overlapp and give us full strand of interest. Remember! We didn't know what the DNA seq was to begin with, it can be some sort of Virus or other microbe but we don't know which!

  • @alexjones1935
    @alexjones1935 4 роки тому +10

    Great video, fast and detailed!

  • @SB-pk8gw
    @SB-pk8gw 10 місяців тому +2

    I have a question: Why is a copy done before the Bridge Amplification? Isn't the Bridge Amplification enough for amplifying my DNA?

  • @tewjiawey4846
    @tewjiawey4846 3 роки тому +1

    Thank you for the video. This made my concept much clear now!

  • @fevroniadermentzoglou8232
    @fevroniadermentzoglou8232 22 дні тому +1

    perfect explanation, thank you

  • @helloisypereira7714
    @helloisypereira7714 2 роки тому +1

    Amazing explanation and video! Thank you.

  • @tark5098
    @tark5098 3 роки тому +2

    thank you for your clear explanation

  • @IcarusLife
    @IcarusLife 3 роки тому +2

    This is the best explanation✨

  • @tathagatabhowmik3053
    @tathagatabhowmik3053 4 роки тому +2

    Very well defined, To the point. Thank you.

  • @reyesbecerraperez3469
    @reyesbecerraperez3469 3 роки тому +3

    Thank you! This was very helpful :)

  • @rdowlati
    @rdowlati Рік тому

    Very well explained. Please also explain depth of read and coverage

  • @danielpintard7382
    @danielpintard7382 Рік тому

    1:54 what was the point of amplifying the dna fragment if the complementary strand is denatured and then washed away?

  • @verdasney
    @verdasney 3 роки тому +4

    Very short and very understandable! Thank you for good work!

  • @carlosbraun5791
    @carlosbraun5791 4 роки тому +3

    Thank you for this vid, but in the compnay video they mentioned an index 1 and idex 2. What are those and which is their purpose????

    • @weege5.45
      @weege5.45 3 роки тому +1

      Paired ends. When doing an alignment one is chemically specified to left side and one to the right. It labels the 5' and 3' ends appropriately for alignment sequencing with something like SPAdes.

  • @lucaskellar4892
    @lucaskellar4892 3 роки тому +2

    very well explained, thank you

  • @quebrachal
    @quebrachal 7 місяців тому

    Hi! This video is great but I still struggle to understand the process. At 01:29, we have a single-stranded forward fragment which is hybridized to the blue oligos and then PCR occurs and the fragment is amplified. In this step, is not happening the same with the reverse fragments, which are hybridized to the red oligos and amplified? What changes in the second step with the bridge building, why is it necessary? I'd really appreciate if someone can help me understand! 🙂

  • @yayahwinarti95
    @yayahwinarti95 Рік тому +1

    Thanks a lot for this incredible explanation. Anw, I have some questions :
    1. What is the purpose of DNA fragmentation? Is there any specific site for this cutting?
    2. Regarding the adaptor, would you mind to explain more about the constituent of this? Is it like primer where we should design it first? How the adaptor can recognize the end of the fragmented DNA?

    • @keb_in
      @keb_in Рік тому +1

      The DNA is fragmented to enhance the speed of the sequencing (it's faster to sequence a lot of tiny bits, than a really long chain), and to reduce the "mistake" rate of the polymerase. plus, the polymerase can fall off if the sequence is extremely long

    • @NahlaElSherif
      @NahlaElSherif Рік тому

      doesnt shorter reads increase the error rate?@@keb_in

  • @DevrelHarry
    @DevrelHarry 3 роки тому +1

    It is great indeed. One question, why do we need to pcr amplify a single strand into many copies in a same spot? At the end isn't it just sequencing a single strand on a particular spot?

    • @henrikslab
      @henrikslab  3 роки тому +3

      PCR is required to get enough material for sequencing. Single strands are not sufficient.

    • @DevrelHarry
      @DevrelHarry 3 роки тому

      @@henrikslab during sequencing reaction after each nucleotide is incorporated, the terminator is removed, as well the fluorescence molecule. Why 1000 to 5000 copies of a single strand is required in the same spot or cluster?

  • @awaiskhan9738
    @awaiskhan9738 Рік тому +1

    I have 1 confusion!
    A single strand has adapter A on one side and adapter B on the other side or end! When it is added to flow cell and its adapter A side binds to its complementary oligonucleotide and DNA generates a complete strand, then the orginal strand is denatured and wash away. Now the strand which is left stand! How can it form a bridge when it is not carrying the sequence of adapter B but its complementary sequence?

  • @hifza2107
    @hifza2107 6 місяців тому

    What is NGS sequencing?
    For that what we do first is fragment our DNA.
    Each fragment will then be attached to two adapters, a side chain nucleotide binding sequence and an oligonucleotide that will bind to the flow cell.
    The DNA is denatured and the fragments then attach to the flow cell. Here DNA amplification by PCR results. The unattached atrand is washed away.
    Hence, bridge amplification is carried out where the strand attached via both it's ends to the flow cell and the PCR is carried out. In this way, both the strands are attached to the flow cell. After several rounds we have the amplified DNA. Having both forward and reverse strands.
    The reverse strands are washed away and the forward strands undergo sequencing by synthesis
    Here each strand has primer attached and one by one a fluorescently labeled nucleotide attaches to the DNA, which is detected each time a nucleotide is incorporated, once the DNA fragments are replicated and nucleotide for one particular strand is detected along with for other strands, bioinformatics tools are used to determine the sequence of entire fragments by placing the sequences side by side and determining the sequence overlaps. The sequence are compared to reference genome

  • @tanatswamashamba9798
    @tanatswamashamba9798 3 роки тому +1

    I'm liking the Next Generation Sequencing (illumina)

  • @89lilly
    @89lilly 3 роки тому +3

    Amazing explanation 🤩🤩🤩 thank you so much!

  • @tamilbiology5078
    @tamilbiology5078 3 роки тому +1

    Superb explanation

  • @msbgoku
    @msbgoku Рік тому +1

    really great helpful explanation thank you much

  • @MrHowtowastetime
    @MrHowtowastetime 3 роки тому +2

    Super clear. Thank you.

  • @ShivendraGupta-xv2zf
    @ShivendraGupta-xv2zf 8 місяців тому

    I have a doubt. Bridge building occurs with both forward (5' to 3') and reverse sequences (3' to 5') right? So when, a bridge is formed by reverse sequence, the amplification of forward sequence is continuous and when a bridge is formed by forward sequence (5' to 3'), the amplification of the reverse sequence is discontinuous (3' to 5') as we see in traditional DNA replication by the formation of Okazaki fragments?

  • @mariela6398
    @mariela6398 4 роки тому +2

    Very well explained. Thanks!

  • @김성경-s7o
    @김성경-s7o 7 місяців тому +1

    I’m high school student..
    How is DNA synthesized in the 3 to 5 direction in the reverse strand?
    Please let me know about reverse strand!!

  • @HewieOG
    @HewieOG Рік тому +2

    amazing tutorial, so well explained, made a very complex subject very simple :)

  • @martavenneri2057
    @martavenneri2057 5 місяців тому +1

    THANK. YOU. SO. MUCH. Really clear

  • @inezodhiambo9057
    @inezodhiambo9057 Рік тому +1

    Thumbs up man, it's incredible. Help more.

  • @documents7840
    @documents7840 2 роки тому +1

    Thank you so much for sharing. It really helped.

  • @aliqassem7780
    @aliqassem7780 3 роки тому +2

    terrific and simple explanation, thanx a lot

  • @en6866
    @en6866 4 роки тому +1

    Very good explanation. Thank you

  • @musakhanmedicalenginee890
    @musakhanmedicalenginee890 11 місяців тому +2

    If we sequence it for the first time,and we will have no reference genome,then what will do? Plzz must reply me

    • @henrikslab
      @henrikslab  11 місяців тому +1

      Don't know the exact case scenario... but in case you sequence cells of an animal such as "Fish X", you may use the reference genome from the most closely related animal of which the sequence is known/published!

  • @gabrielphilips6980
    @gabrielphilips6980 Рік тому +2

    How do you manipulate DNA? I suppose you cannot grab them using tweezers, right?

    • @henrikslab
      @henrikslab  Рік тому +1

      There are multiple ways to do so. But in many cases the "tweezers" scientists use are enzymes, that bind to specific or in other cases more unspecific DNA and can delete (cut out) base pairs or even add (insert) new ones. A cool example of how to manipulate DNA is the CRISPR Cas9 system...
      See here: ua-cam.com/video/4knIPgD6h1U/v-deo.html

  • @amirarayanerighi2706
    @amirarayanerighi2706 2 роки тому +1

    very clear explanation thank you very much

  • @mariangeladagati5260
    @mariangeladagati5260 11 місяців тому

    This is the greatest explanation I have seen about the procedure. Kudos!

  • @angelhimelisa
    @angelhimelisa Рік тому +3

    I feel stupid cuz everyone's like.. this is the best explanation I've ever found! And I *still* don't know what the hell he's talking about im gonna fail rofl

  • @mariannamycroft2611
    @mariannamycroft2611 3 роки тому +2

    You're a saviour! Thanks

  • @yuqingcai3447
    @yuqingcai3447 Рік тому +1

    Thank you so much sir.... You are my savior!

  • @ammarahchaudhary2820
    @ammarahchaudhary2820 4 роки тому +1

    Straight to the point
    Use good English with good explanation .
    Thanx✌👍👏

  • @giorgiafiorino3920
    @giorgiafiorino3920 10 місяців тому +1

    Thanks so much, really well done!

  • @debanjanchowdhury4397
    @debanjanchowdhury4397 8 місяців тому +1

    Well explained 👏

  • @marianalamuno9445
    @marianalamuno9445 4 роки тому +2

    Great video! very clear thank you

  • @rachaelgoh4310
    @rachaelgoh4310 3 місяці тому

    Wanted to ask why do you wash away all of the reverse strands after PCR? Does it matter whether you wash away the reverse or forward strand?

  • @Celtjak7
    @Celtjak7 8 місяців тому +1

    So in order to 'see' the DNA, you need to see the fluorescence, so you need the leading strand to be fluorescent, therefore needing copies of the template strand.

  • @henrikslab
    @henrikslab  4 роки тому +1

    Which topics should I cover in my next videos? Make suggestions below this comment and like the ones you are most interest in

    • @SK-pi6hr
      @SK-pi6hr 3 роки тому

      Gene cloning, micro array, protein assay

    • @henrikslab
      @henrikslab  3 роки тому

      @@SK-pi6hr Do you mean Western Blot with Protein Assay? Or do you mean some concentration determination of protein (e.g. Bradford Assay)

    • @SK-pi6hr
      @SK-pi6hr 3 роки тому +1

      @@henrikslab concentration determination of protein

    • @henrikslab
      @henrikslab  3 роки тому

      @@SK-pi6hr Your wish is granted! Upload: Today :)

  • @daoyang491
    @daoyang491 3 роки тому +1

    I love the explanation

  • @gildashounmanou4301
    @gildashounmanou4301 4 роки тому +2

    straight to the point

  • @hudaalhajj8017
    @hudaalhajj8017 7 місяців тому +2

    very good. now you can go to my exam and maybe pass

  • @isadorah4969
    @isadorah4969 7 місяців тому

    Does the fluorescence of the paired nucleotides goes away when it pairs to the DNA? I mean, if the sequence comes to be yellow-blue-yellow-blue as in TATA boxes for example, would it emit green light because of the combination of the colors?

  • @yogitabasnal
    @yogitabasnal 3 роки тому

    Adopter is for binding of primer right?? I think there is one more use of adopter. I wanna know about adopter please

  • @rimelhajj6520
    @rimelhajj6520 3 роки тому +2

    You saved me! Thankyou so muchh😭😭❤️

  • @arnemuller4629
    @arnemuller4629 4 роки тому +2

    Hey, I´d like to use some screenshots of your graphical illustration in a powerpoint presentation, if that´s okay for you (master student seminar, im a student)? - really liked your video, it made understanding Illumina much easier! Thank you so much

  • @sienablier8931
    @sienablier8931 2 роки тому

    Thank you so much, this was super well explained!

  • @ZLYang
    @ZLYang Рік тому

    Thank you! Clear explanation.

  • @faramund9865
    @faramund9865 3 роки тому +1

    Just a request there Henrik. It'd be lovely if there'd be fewer rare, field specific, Latin/Greek/French words in these kinds of videos. Makes it really hard to understand for a layman. So I would really love if you could try and translate these into common Germanic words that everyone knows.
    Examples: Ligation, DNA Adaptors, sequence, complimentary, hybridize, denaturation, polymirazed chain reaction, synthesized, olegol? nucleotides (genuinely don't know what you said there). I could go on and on. I hope you get the gist of what I'm saying. Would really help a lot of people understand what's going on much more easily.

    • @henrikslab
      @henrikslab  3 роки тому +1

      Thank you for that kind of nice feedback! Yes, I definitely agree with you - I guess the videos are not really designed for a pure layman... most people watching the videos are biology students themselves (so I honestly kind of expect them to know)
      Nevertheless, I take this point and might think twice before using a specific term. Thanks!

    • @faramund9865
      @faramund9865 3 роки тому

      @@henrikslab Thanks for your response and for considering my advice!

  • @Pauluna123
    @Pauluna123 4 роки тому +1

    Nice well explained lesson