454 Sequencing

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  • เผยแพร่เมื่อ 11 ม.ค. 2025

ความคิดเห็น • 141

  • @willkerry
    @willkerry 8 ปีที่แล้ว +59

    Good video but your explanation of the emulsion PCR step is a bit off. You seem to suggest the strand complementary to the original template (created by extension of the bead oligo) breaking off after denaturation and annealing to another one of the bead oligos. This is not what happens as 1.) this strand is attached to the bead and therefore cannot move and 2.) it wouldn't bind to the bead oligos as it is not complementary, it's identical. Instead, after the first denaturation step it is the original template that breaks off and binds to another bead oligo (as before) and again a complementary strand is created. At the same time, a PCR primer will bind to the 5' end of the first complementary strand (the end away from the bead) and will be extended towards the bead to generate another copy of the original template strand. This process is repeated until the bead is covered in clonally amplified copies of the original template.

    • @DileepKumar-tj9ck
      @DileepKumar-tj9ck 7 ปีที่แล้ว

      Thank you, William Kerry.

    • @hashh555
      @hashh555 7 ปีที่แล้ว +1

      Was searching for this explanation and couldn't find - indeed makes sense, thank you.

    • @dadsfriendlyrobotcompany
      @dadsfriendlyrobotcompany 6 ปีที่แล้ว +1

      I was thinking this exact thing as he was explaining it

    • @Thomaaasooo
      @Thomaaasooo 5 ปีที่แล้ว

      @Q Suraiya you are correct

    • @goliathustcg5952
      @goliathustcg5952 4 ปีที่แล้ว +2

      Thanks. I'm wondering: How exactly do you make sure only one DNA sequence lands on a bead, without others landing there at the same time and the bead therefore being incosistant?

  • @Jonathan544200
    @Jonathan544200 7 ปีที่แล้ว +1

    Brother, i will be expected to present and explain a machine from a company which i utterly want them to hire me, i decided to present 454 sequencing and this video saved my life. thanks from turkey

  • @MoOnassKA
    @MoOnassKA 7 ปีที่แล้ว +2

    You're the best , I can't undersand in my class, and you make things so easy to understand , thank you !

  • @rcr8135
    @rcr8135 3 ปีที่แล้ว +2

    Thank you Shomu sir, your way of explaining concepts is excellent.

  • @oznurmete6685
    @oznurmete6685 8 ปีที่แล้ว

    Thank you with my all heart. i watched almost all of your videos. please know there is a person who pray for you :)

  • @benda-annevanniekerk6833
    @benda-annevanniekerk6833 8 ปีที่แล้ว +2

    Helped me to actually make sense of the textbook, thank you!

  • @macplay9765
    @macplay9765 7 ปีที่แล้ว +2

    Sir ji you're great ... u provide exactly those material which we want.. hats off.. keep doing it...

  • @dennislin303
    @dennislin303 6 ปีที่แล้ว +1

    Very well explained especially for a beginner like myself. Thanks for sharing!

  • @amandinevivet9233
    @amandinevivet9233 ปีที่แล้ว +1

    Your video is perfect ! I understood everything, thank you very much :)

  • @fatimabatool4010
    @fatimabatool4010 8 ปีที่แล้ว

    Thank you so much Sir. It helped me so much. The way you explain is so understandable for everyone. Thanks a bunch. God bless you

  • @lenkaobrtancova6949
    @lenkaobrtancova6949 7 ปีที่แล้ว +4

    well done prof! thank you for this great explanation :)

  • @tomasvalek381
    @tomasvalek381 2 ปีที่แล้ว +1

    Very nice explanation, I learn a lot. THX

  • @mariamalmahdy6372
    @mariamalmahdy6372 8 ปีที่แล้ว

    mashaAllah very nice and helpful explanation thank u so much u r genius really though you look very young , may Allah bless you ... I watch you lectures since last year and thy ve been always a great help to me .. thank you dear

  • @PRAVEENDEP
    @PRAVEENDEP 8 ปีที่แล้ว +2

    Excellent explanation !! your every lecture's video is so nicely explained, Good to helps other students or researchers ! Good luck for future !! Go ahead....... :)

  • @isaurasilva4103
    @isaurasilva4103 7 ปีที่แล้ว +2

    no hablo ingles pero entiendo mas o menos y usted lo hizo tan sencillo, que comprendí muy bien, muchas gracias :D

  • @itzelelizalde4758
    @itzelelizalde4758 8 ปีที่แล้ว +1

    Thank you! From México.

  • @alexstar7995
    @alexstar7995 8 ปีที่แล้ว

    Thank you for the indepth explanation, appreciated it.

  • @juliabl6471
    @juliabl6471 7 ปีที่แล้ว

    at minute 5:57 why is it necessary to use adaptor B if it is not useful to fix the DNA to the particle?

  • @coreyman7634
    @coreyman7634 8 ปีที่แล้ว

    Thank you, these are great videos!!!

  • @snehalchavda5738
    @snehalchavda5738 2 ปีที่แล้ว +1

    Very informative

  • @gurkamalkaur9262
    @gurkamalkaur9262 7 ปีที่แล้ว

    You r the best 👍🏻 keep it up

  • @assistantprofessor8049
    @assistantprofessor8049 8 ปีที่แล้ว +1

    Thank you for informative video but am confused with primer addition at the end. As I know first we use primer with adaptor sequences to make copies of complementary DNA stand by pcr and then run on 454 sequencer. But in your video you explain that we don't need pcr to make copies. Then how we will add primer and adaptor sequence to target dna for sequencing. Please if possible make it more clear for me and please also draw a schematic work flow of this whole process in Sequence from 1 to end. Thank you you did very job for us. GOD BLESS YOU MORE SUCCESS

  • @melissabouzar5273
    @melissabouzar5273 8 ปีที่แล้ว

    Thank you! you are saving me !

  • @botanica19
    @botanica19 ปีที่แล้ว +1

    Well presented...

  • @rumeysadgn
    @rumeysadgn 8 ปีที่แล้ว

    So many thanks. It is really helpful. :)

  • @jurgenpenn4624
    @jurgenpenn4624 8 ปีที่แล้ว +1

    Great explanation, thanks.

  • @tetsu6strings
    @tetsu6strings 8 ปีที่แล้ว

    Thanks for the great video!

  • @Mechee14
    @Mechee14 7 ปีที่แล้ว +4

    hello! i have a question, isn't 454 same as pyrosequencing? becouse here you explain like each nucleotide has a flourescent tag, but pyro uses luciferase....

    • @anjalishukla61
      @anjalishukla61 7 ปีที่แล้ว

      Meche Landeira even i have the same question and confused in between pyro and 454

    • @bappisarkar850
      @bappisarkar850 7 ปีที่แล้ว

      yahh exactly... please rectify the explanation.

  • @Drutten87
    @Drutten87 8 ปีที่แล้ว

    Thank you! You're the boss

  • @ummafatema9534
    @ummafatema9534 8 ปีที่แล้ว +1

    Thank you. This video is well understandable. But when the primer is actually added?

    • @fatimabatool4010
      @fatimabatool4010 8 ปีที่แล้ว

      Primer is added after loading the beads into wells.

  • @nitinsaxena7370
    @nitinsaxena7370 8 ปีที่แล้ว

    Thank you -- very well done!!

  • @sayansarkar8546
    @sayansarkar8546 4 ปีที่แล้ว

    Plss make a video about emulsion PCR...
    It will be very helpful

  • @radhekrishnamotivationgang6115
    @radhekrishnamotivationgang6115 2 ปีที่แล้ว +1

    Thank you so much sir

  • @nikmishin9
    @nikmishin9 8 ปีที่แล้ว +5

    Hi Shomu, thanks for great video. As far as I understood the method reads nucleotides in order they actually appear on the strand. What I don't understand is how the method prevents the same nucleotides from different parts of the strand from being read all at once. So if we have the following sequence ATTGCAA and we add Thymine we detect just one fluorescence as T bonded to the first A. How does it not bond to the final AA as well?

    • @shantanusingh8267
      @shantanusingh8267 3 ปีที่แล้ว +1

      I also have the same problem. If you got your answer kindly reply.

  • @ilariaunali7202
    @ilariaunali7202 8 ปีที่แล้ว

    Thank you for your explanation! you're saving me ahah

  • @spandanayadav3549
    @spandanayadav3549 6 ปีที่แล้ว

    Good vedios sir very informative.but the only problem is with sound please update sound system .thank you

  • @zainabalalwani3170
    @zainabalalwani3170 3 ปีที่แล้ว +1

    Excellent

  • @sdfPZXC
    @sdfPZXC 8 ปีที่แล้ว

    great explanation, Thanks XD

    • @sdfPZXC
      @sdfPZXC 8 ปีที่แล้ว

      would be cool to know how fluorescent was generated.

    • @sdfPZXC
      @sdfPZXC 8 ปีที่แล้ว

      oh nevermind it in a pyrosequencing vdo XD

  • @omedhasan6332
    @omedhasan6332 9 ปีที่แล้ว +1

    well done brother

  • @omaraalwahab640
    @omaraalwahab640 8 ปีที่แล้ว

    Thank u very much 😍

  • @subhalaxmisahu1886
    @subhalaxmisahu1886 6 ปีที่แล้ว +1

    Where is the bead present? Is it in the tube naturally?

  • @jigyasasharma01
    @jigyasasharma01 7 ปีที่แล้ว

    ty for saving my xms :)

  • @bruce012324
    @bruce012324 8 ปีที่แล้ว

    how is 454 different from pyrosequencing? i notice that 454 uses two adapters while pyro uses one, but isn't using one adapter a lot more easier? whats the advantage of 454 and pyro method?

  • @kashafrasul3952
    @kashafrasul3952 4 ปีที่แล้ว

    Kindly give lecture on 3-C, 4-C 5-C, ChIA-PET AND Hi-C TECHNOLOGIES.

  • @khatirsafi5079
    @khatirsafi5079 5 ปีที่แล้ว

    HI sir i have a question is pyro sequencing and 454 sequencing are same if not ?AND why we use the second beads on the other end of template dna

  • @quentin-v9d
    @quentin-v9d 8 ปีที่แล้ว +1

    Hey man, I like your video its helped me out and is very clear. The only bit I'm confused about it the end. So you have all these fragments (I'm assuming there are thousands of fragments) of double stranded DNA composed of the target single strand and the fluorescent single strand from which we derived the target sequence. I don't understand how these fragments are put together into the correct order of the actual full length DNA molecule of whatever we sampled?

  • @ramonmassoni9657
    @ramonmassoni9657 8 ปีที่แล้ว

    Awesome bro, you are a genius! One question, is the release of pyorphosphate after each nucleotide is added the way by which the fluorescence is detected?

  • @4BFF9
    @4BFF9 8 ปีที่แล้ว

    perfect, thank you! :)

  • @TS-dp2nv
    @TS-dp2nv 8 ปีที่แล้ว

    Brilliant!

  • @sumenmajumder9900
    @sumenmajumder9900 8 ปีที่แล้ว

    I am confused. How target strand is used as complementary strand in sequencer? I think complementary strand ( produced in beads) is used as complementary strand in sequencer.

  • @jurgenstuderegger4972
    @jurgenstuderegger4972 6 ปีที่แล้ว +1

    Hey! Thanks for uploading those videos. I'm a bit confused. Is this really the 454 sequencing? 454 is the same as pyrosequencing which uses luciferase, or am I wrong? This video looks like the cycle-sequencing. In any case, thank you!

  • @monzurmorshed.
    @monzurmorshed. 5 ปีที่แล้ว +1

    Thank you!

  • @leilasaadi5952
    @leilasaadi5952 2 ปีที่แล้ว

    How do we know which dna fragment (on the bead) the light belongs to?

  • @lukash.8788
    @lukash.8788 2 ปีที่แล้ว

    So it’s basically ion sequencing but instead of protonlevels you just measure the fluorescence?

  • @subhalaxmisahu1886
    @subhalaxmisahu1886 6 ปีที่แล้ว +5

    Why it is called as 454 sequencing?

    • @divyanshutiwari9916
      @divyanshutiwari9916 5 ปีที่แล้ว +3

      @subhalaxmi sahu
      454 life science is the name of company that has given the technique...thats why it is said so

  • @raquelandreaa
    @raquelandreaa 6 ปีที่แล้ว +1

    Thank you!!

  • @128scarface
    @128scarface 9 ปีที่แล้ว

    Thanks for the video. So, how do the adapters bind to the dsDNA? is it just a random event?

  • @harshitakashyap2014
    @harshitakashyap2014 3 ปีที่แล้ว

    why it has got the name as "454" sequencing? Is there any significance to this number?

  • @muhammedyousafk4115
    @muhammedyousafk4115 2 ปีที่แล้ว +1

    Thankyou sir

  • @lollsazz
    @lollsazz 8 ปีที่แล้ว +20

    Only ONE fragmentof DNA is caught per bead - otherwise this wouldn't make sense

  • @Jajati32
    @Jajati32 5 ปีที่แล้ว

    I think above explained method is ABi SOLID sequencing method..Not the 454 sequencing method..

  • @faiqaatique534
    @faiqaatique534 6 ปีที่แล้ว

    Well done

  • @umaraziz335
    @umaraziz335 3 ปีที่แล้ว

    hi sir hope you will be fine ,why 454 sequence is called ,454 ????? please explain it.

  • @johannesmthembu8663
    @johannesmthembu8663 7 ปีที่แล้ว

    wait i wanna know something Dr Shomu there is a video that u uploaded by the tittle "pyrosequencing" and this one of "454 sequencing"...is there a difference between pyrosequencing and 454 sequencing since i feel as if what u have explained is the same for both videos ...please help.thank u

  • @venkataadityaganta1967
    @venkataadityaganta1967 4 ปีที่แล้ว

    what is the special purpose of using bead. Please someone answer and clarify to me that single point.

  • @IAM-ii4nq
    @IAM-ii4nq 7 ปีที่แล้ว

    Thanks Prof

  • @bszoologest3303
    @bszoologest3303 4 ปีที่แล้ว +1

    sir what is the difference between pyrosequencing and 454 sequencing

  • @simplesimi798
    @simplesimi798 6 ปีที่แล้ว +1

    Thank you

  • @aishasaleem3738
    @aishasaleem3738 7 ปีที่แล้ว +1

    very well explanation sir

  • @swapnikasharma7104
    @swapnikasharma7104 4 ปีที่แล้ว +1

    Thanks

  • @sportsgaming2548
    @sportsgaming2548 4 ปีที่แล้ว

    Sir is it same as Solid D sequnc?

  • @tanishadas-mu4qi
    @tanishadas-mu4qi ปีที่แล้ว

    Sir please make a vedio on signature sequences

  • @believeinyourself7947
    @believeinyourself7947 2 ปีที่แล้ว

    Sor kindly suggest a book for ngs study

  • @9804871393
    @9804871393 7 ปีที่แล้ว

    as per my knowledge 454 and pyro are same sequencing method.

  • @nailaimtiaz3617
    @nailaimtiaz3617 3 ปีที่แล้ว

    Plz make video for SOLiD sequencing

  • @laul3617
    @laul3617 8 ปีที่แล้ว

    Muchas gracias!!

  • @ReplicatedMirror
    @ReplicatedMirror 3 ปีที่แล้ว

    I am a little bit confused that which strand was going where and which was target strand. Just messy man...
    please, make another clear concept video,please.

  • @vancewade6251
    @vancewade6251 8 ปีที่แล้ว

    I'm curious to know how the machine inputs one type of dNTP at a time and then washes it and adds the next one so damn quickly

  • @dc33333
    @dc33333 6 ปีที่แล้ว

    Video not accurate.The statement there is no PCR is not correct; emulsion PCR. Other inaccuracies as noted below

  • @abhijitpaul8509
    @abhijitpaul8509 6 ปีที่แล้ว +1

    why it is called 454 sequence?

    • @rishianand100
      @rishianand100 5 ปีที่แล้ว

      454 is the name of the company now called Roche/454

  • @Maitedmm
    @Maitedmm 8 ปีที่แล้ว

    THANKS!!

  • @noorajmal62
    @noorajmal62 7 หลายเดือนก่อน

    Why 454 sequence is called 454 sequence??

  • @omedhasan6332
    @omedhasan6332 9 ปีที่แล้ว +2

    really i love you

  • @saraha6980
    @saraha6980 3 ปีที่แล้ว

    Thank you very much for helping me in every single exam (✦థ ェ థ)

  • @ricardosantanagalego7938
    @ricardosantanagalego7938 7 ปีที่แล้ว

    Muito very good

  • @johnydangi8599
    @johnydangi8599 9 ปีที่แล้ว

    must h....

  • @siriveludharani6338
    @siriveludharani6338 3 ปีที่แล้ว

    🙏🙏🙏🙏🙏🙏🙏🙏🙏🙏🙏

  • @leonardomohamed5512
    @leonardomohamed5512 8 ปีที่แล้ว

    You have been more celar thad mi

  • @akankshabharti7013
    @akankshabharti7013 7 ปีที่แล้ว

    confusing

  • @joshuaspall
    @joshuaspall 7 ปีที่แล้ว

    dude go inside

  • @fahlihamza7937
    @fahlihamza7937 8 ปีที่แล้ว

    send