454 Sequencing

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This DNA sequencing lecture explains about the 454 sequencing technology. 454 sequencing is a type of next generation DNA sequencing technology using the DNA probe attached with the insoluble bead.
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Опубликовано:

5 окт 2024

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Комментарии : 142

@willkerry 7 лет назад

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 7 лет назад

Thank you, William Kerry.

@hashh555 7 лет назад

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

@dadsfriendlyrobotcompany 6 лет назад

I was thinking this exact thing as he was explaining it

@Thomaaasooo 5 лет назад

@Q Suraiya you are correct

@goliathustcg5952 3 года назад

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?

@benda-annevanniekerk6833 8 лет назад

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

@rcr8135 2 года назад

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

@shomusbiologyofficial 2 года назад

Thank you so much for appreciating my efforts

@MoOnassKA 7 лет назад

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

@lenkaobrtancova6949 6 лет назад

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

@oznurmete6685 8 лет назад

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

@dennislin303 5 лет назад

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

@shomusbiologyofficial 5 лет назад

Glad to hear that you are getting benefit from

@Jonathan544200 7 лет назад

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

@tomasvalek381 2 года назад

Very nice explanation, I learn a lot. THX

@shomusbiologyofficial 2 года назад

You're welcome

@amandinevivet9233 Год назад

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

@shomusbiologyofficial Год назад

You're welcome

@fatimabatool4010 7 лет назад

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

@shomusbiologyofficial 7 лет назад

+Fatima Batool that's great. I am glad that it helped you

@macplay9765 7 лет назад

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

@assistantprofessor8049 7 лет назад

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

@isaurasilva4103 7 лет назад

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

@itzelelizalde4758 8 лет назад

Thank you! From México.

@PRAVEENDEP 8 лет назад

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

@shomusbiologyofficial 8 лет назад

+Praveen Prajapat thank you. Glad you liked my lectures

@ummafatema9534 8 лет назад

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

@fatimabatool4010 7 лет назад

Primer is added after loading the beads into wells.

@snehalchavda5738 2 года назад

Very informative

@shomusbiologyofficial 2 года назад

Thank you

@botanica19 8 месяцев назад

Well presented...

@shomusbiologyofficial 8 месяцев назад

Thank you

@nikmishin9 8 лет назад

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 3 года назад

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

@radhekrishnamotivationgang6115 2 года назад

Thank you so much sir

@shomusbiologyofficial 2 года назад

You're welcome

@mariamalmahdy6372 8 лет назад

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

@alexstar7995 7 лет назад

Thank you for the indepth explanation, appreciated it.

@shomusbiologyofficial 7 лет назад

+Alex Star thank you. Glad you liked my lectures

@sayansarkar8546 4 года назад

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

@shomusbiologyofficial 4 года назад

Okay

@zainabalalwani3170 2 года назад

Excellent

@shomusbiologyofficial 2 года назад

Thank you

@subhalaxmisahu1886 6 лет назад

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

@spandanayadav3549 6 лет назад

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

@jurgenpenn4624 8 лет назад

Great explanation, thanks.

@shomusbiologyofficial 8 лет назад

+Jürgen Penn thank you.

@Mechee14 7 лет назад

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 6 лет назад

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

@bappisarkar850 6 лет назад

yahh exactly... please rectify the explanation.

@subhalaxmisahu1886 6 лет назад

Why it is called as 454 sequencing?

@divyanshutiwari9916 5 лет назад

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

@quentin-v9d 8 лет назад

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?

@melissabouzar5273 8 лет назад

Thank you! you are saving me !

@muhammedyousafk4115 2 года назад

Thankyou sir

@shomusbiologyofficial 2 года назад

You're welcome

@leilasaadi5952 Год назад

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

@kashafrasul3952 4 года назад

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

@coreyman7634 8 лет назад

Thank you, these are great videos!!!

@monzurmorshed. 5 лет назад

Thank you!

@shomusbiologyofficial 5 лет назад

You're welcome

@lukash.8788 2 года назад

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

@bruce012324 7 лет назад

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?

@ramonmassoni9657 7 лет назад

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?

@sumenmajumder9900 8 лет назад

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.

@tetsu6strings 8 лет назад

Thanks for the great video!

@swapnikasharma7104 4 года назад

Thanks

@shomusbiologyofficial 4 года назад

You're welcome

@umaraziz335 2 года назад

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

@juliabl6471 7 лет назад

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

@harshitakashyap2014 3 года назад

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

@tanishadas-mu4qi 9 месяцев назад

Sir please make a vedio on signature sequences

@shomusbiologyofficial 9 месяцев назад

Okay

@gurkamalkaur9262 7 лет назад

You r the best 👍🏻 keep it up

@Jajati32 4 года назад

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

@nitinsaxena7370 8 лет назад

Thank you -- very well done!!

@shomusbiologyofficial 8 лет назад

+Nitin Saxena thank you very much

@believeinyourself7947 2 года назад

Sor kindly suggest a book for ngs study

@omedhasan6332 8 лет назад

well done brother

@shomusbiologyofficial 8 лет назад

Thank you.

@raquelandreaa 6 лет назад

Thank you!!

@shomusbiologyofficial 6 лет назад

You're welcome

@venkataadityaganta1967 4 года назад

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

@simplesimi798 6 лет назад

Thank you

@shomusbiologyofficial 6 лет назад

You're welcome

@jurgenstuderegger4972 6 лет назад

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!

@sportsgaming2548 4 года назад

No its differ

@rumeysadgn 7 лет назад

So many thanks. It is really helpful. :)

@shomusbiologyofficial 7 лет назад

+Rumeysa Doğan thank you. Glad you liked my lectures

@theworld3531 5 лет назад

is the sequence like CGGGAA or cGA

@IAM-ii4nq 7 лет назад

Thanks Prof

@omaraalwahab640 7 лет назад

Thank u very much 😍

@nailaimtiaz3617 2 года назад

Plz make video for SOLiD sequencing

@shomusbiologyofficial 2 года назад

Okay

@TS-dp2nv 8 лет назад

Brilliant!

@jigyasasharma01 6 лет назад

ty for saving my xms :)

@khatirsafi5079 5 лет назад

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

@faiqaatique534 6 лет назад

Well done

@sportsgaming2548 4 года назад

Sir is it same as Solid D sequnc?

@laul3617 8 лет назад

Muchas gracias!!

@lollsazz 7 лет назад

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

@sailajakambhampati9071 3 года назад

so what are the software programs involved in this?

@bszoologest3303 3 года назад

sir what is the difference between pyrosequencing and 454 sequencing

@bszoologest3303 3 года назад

please explain sir

@bszoologest3303 3 года назад

are they same ???

@Drutten87 7 лет назад

Thank you! You're the boss

@ilariaunali7202 7 лет назад

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

@johannesmthembu8663 7 лет назад

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

@noorajmal62 4 месяца назад

Why 454 sequence is called 454 sequence??

@ReplicatedMirror 3 года назад

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.