Illumina sequencing | DNA sequencing by synthesis
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- Опубліковано 15 вер 2024
- Illumina sequencing by synthesis - illumina sequencing process is explained in this video lecture. Illumina is one type of second generation DNA sequencing technique to get the sequence if target DNA easily and fast.
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The “sequencing-by using-synthesis” science now utilized by Illumina used to be at first developed by using Shankar Balasubramanian and David Klenerman at the college of Cambridge. They headquartered the company Solexa in 1998 to commercialize their sequencing procedure. Illumina went on to purchase Solexa in 2007 and has built upon, and speedily accelerated the long-established technology.
Illumina sequencers are presently probably the most generally used sequencing platform within the subsequent-new release sequencing (NGS) field. Illumina uses drift-cellphone floor for clustering DNA by way of ‘bridging amplification’, which generates clusters with hundreds of thousands of identical, single-stranded (ss), floor-hooked up DNA molecules After primer annealing, fluorescently labeled dATP, dGTP, dCTP and dTTP are introduced to the three′ end of the primer in line with the complementary base of the template strand. The fluorescently labeled nucleotides are chemically blanketed at the 3′ hydroxyl staff, which prevents the addition of greater than a single nucleotide per cycle. The digital camera then takes a snapshot of the go with the flow cellphone to realize the fluorescence from the final incorporated nucleotide of each cluster. The 3′ hydroxyl protection group as well as the fluorophore is enzymatically cleaved to proceed to the following cycle of the sequencing reaction. This stepwise addition of sequencing reactions is fascinating when sequencing homopolymer (repeating stretch of 1 form of nucleotide), which is often complicated for different sequencing platforms147. Moreover, the throughput of Illumina sequencers per sequencing run is 10-one hundred instances bigger than that of other sequencing platforms. Paired-end sequencing capabilities are additionally well headquartered, and these can make amends for the shorter learn length and present extended accuracy by studying the equal DNA template twice. The capabilities problem of Illumina sequencing technology is that the buildup of uncleaved fluorophores or protection agencies from each and every step can set off excessive noise and expand substitution error within the later sequencing cycles.
The so called professors or doctors at universities are just doing the job for living, not coz they have passion for science. Therefore, they always fail to communicate science in a simple way as you do brother. Much appreciation! you are making huge difference.
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I am doing MSc Biotechnology in UK but still i look up to your videos which makes easier also time saving .I hope one day i would make you proud sir 🌻Thank you so much for being an inspiration
Glad to help you
I am just a High school student learning this😅
Shomus Biology Way Beyond His Time What Other Institutes are teaching today that was already taught way back in 2015 😮
I really don't have sufficient words to express how grateful I am! you have made biochemistry and molecular biology easier to learn. thank you so much.
thank u sir when i am unable to understand any topic i opened your videos and and i get solution of my problem there .you are very intelligent and way of teaching is mindblowing
+Umer Khan thank you. I am glad that you are getting benefit from the videos
Can you make a video on epigenetics, bisulfite conversion, methylation-specific PCR, and bisulfite sequencing?
Thanks!
thank you for this video, it is the best explanation of Illumina sequencing we can get!!!
Thank you for the excellent explanation of the bridge amplification method.
Thank You so much bro... You are so good all the way... Proud to be Bioinformatitian... Love You...
The way you explain these things, it makes it easy to understand
Thank you so much for making concepts understandable.
Glad to hear that you're getting benefit from my lectures
Awesome explanation
Thank you
Thankyou Shomu this is the best explanation :)
+Humairah Rashad you're welcome
thank u sooooooo much sir......for this vidio .it is really very helpful to me
Thank u very much sir . U made this topic very easy to understand.
Thank you Shomu ! Excellent explanation, so helpful as always :D
WHY DO WE FRGAMENT D DNA BEFORE SEQUENCING THEM? AND CAN WE ALSO USE SONICATION METHOD TO DO SO ?
There is limit to reading the base pairs . No machine can read it all once
the primordial shomuji
Very well explained, thanks
You're welcome
Excellent explanation, thank you shomu
Thank You! ❤
You're welcome
very good explaination sir..
In the bridge dissociation step....what happens to the stranded DNA that it dissociates and becomes single stranded? and secondly...Why one end of the strand remains attached while the other is free? Why don't both the ends tend to become free altogether? Kindly explain it...thanks!!!
Sir is there any recommended book to read sequencing?
Thank you so much for the explanation
Indians are the best!! thanks dude
+Francisco De Francischis you're welcome
SHOMU! you are amazing! keep up the good work
Great explanation👍🏻
Thank you
Thank you so much. Lifesaver!!
+Sylvia Jiménez glad I can help
Loved this video!
Thank you
Thank you sir very clear
You're welcome
you ARE SIMPLY THE BEST
+Vinie Kouamou thank you. Glad you liked my lectures
please keep it up!
Thank u....ur best ❤️
Thank you
Hi, can you explain to me why we need 2 reads when sequencing? Also, is barcoding necessary when analyzing self "made" (PCR) 16s rRNA gene fragments? or is it only useful when you get samples from libraries?
I'm still trying to figure this all out myself, but for the barcoding question, I think it's only necessary for high throughput or when you don't know exactly what you have. I don't know what you mean by self made, but if you clone a 16s fragment for identification you can send it for sanger sequencing which is much cheaper
ThankYou.
You're welcome
Sir can you upload a vedio regarding the relaxation of sequnece reads in bioinformatics?
how does the denaturing ensure that each of the type of ends will be attached to the surface and other end will be free? I mean why not both violet coloured ends be attached and both pink coloured ends be free? why does it have to be one of each? Can you please clarify?
Another great video!
Hello Sir. Thank you
You're welcome
How will be the loci which have 50% probability to have two of any base (like you explained) be resolved? stated other way round, is that a limitation that Illumina have or can it be resolved by resequencing or Gap joining algorithms?
can you please make a video on PacBio sequencing? i couldn't find it on your channel
Well explained
very good
Love you shoumu !
thank u . . . . . for making this.. . . . adil rahim abulwalikhan uni mardan kpk pakistan
+Adil Rahim glad to hear that
hi sir i hope you see this, i am very grateful for your videos they have helped me immensely those past few years. i have a question,i cant grasp why there is an overlapping in the final sequenced product. didn't we fragment the whole genome at the beginning? so what causes those overlaps? thank you so much
Fragmenting genome randomly generates the overlaps
Sehr Gut. vielen dank!
What the different between Next generation and sanger and when i used this two ??
Are different fragments amplified on the same slide?
Why adapters are added on both sides of the dna fragment? Is it for bridge amplification?
Please make the photos clear.. all of your explanations will be based on that pics only.. so that one can easy follow you... And I think little more information should be included on reads...
I have one question... your DNA is sequenced by ilumina miseq is of 5MB size. U have to get 10x coverage . How much data do u need?
does bridge amplification produce a duplicate or a compliment sequce??
now a days, data we will get from company but after getting NGS data (illumina)how to its a big problem, which software we have to yse and how, can you tell?
Does this pcr occurs in iso thermic condition?? Its not like normal pcr
Nice
Thank you
A video on SOLiD sequencing please
Okay
can you please refer a book for studying all type of sequencing method?
What makes the DNA bend ??
Hi Can i please request you to do a video on ion torrent sequencing plz plz
Can i get those images you are using in this video of illumina sequencing???
I didn't get the point once it bend down then how it straightend later
what is the nature of adapters?
Breeege.... iiiis, u dnt understand why do we do in reverse and forward strand and why do u it
Subtitles don't match with what you say
Nothing is visible sir
SOLiD seq plzzzzz
Okay
Anyone from KU?😂