Ion torrent sequencing

[@tsheringdorji4648]

OMG! After watching this, I understood the whole principle of NGS. Explained in a very clear and simple way. Thank you.

[@shomusbiologyofficial]

You're welcome

[@Lineapetrela]

Mr. Shomu you are an excellent teacher, thank you for spreading all your knowledge

[@fleridafigueroa5245]

I cannot even explain how grateful I am, i couldnt understand this untill now. Thanks!

[@Santificated]

Shomu you're awesome man. Thank you so much. I've got an advanced biochemistry test tomorrow and your videos have helped heaps! Keep up with the good work!

[@anirudhrana-bs5sl]

Nice sir

[@javiervalls2021]

In emulsion PCR we have a lot of beads, that are attached to a specific oligo, that allow us to amplify our fragment (which has a sequence we want to know). It is impossible that one bead could be attached to different fragments as you say in minute 12:46, isn't it? In emulsion PCR

[@AzminaAlisa]

Thank you so much Mr Shomu, you are truly the best!

[@shomusbiologyofficial]

Thank you so much for appreciating my efforts

[@caitlinsilva4510]

This is actually incredible, can't believe I'm just now finding this man, this was so helpful. tHANK YOU

[@shomusbiologyofficial]

You're welcome

[@vivianhuang6577]

So glad find this KZbinr! Really benefits me a lot.

[@shomusbiologyofficial]

Thank you. Glad you liked my lectures

[@hs_wears]

I don’t think I can understand it better tha this… thank you for breaking it down👏

[@shomusbiologyofficial]

You're welcome

[@hyuit367]

Amazing job at explaining concepts. Very good job and excellent resource for biology students in post-secondary education.

[@shomusbiologyofficial]

Thank you. Glad you liked my lectures

[@timothyongaba1962]

I lloovveee how passionate you are in all your lectures Shomu😂🤝🏾. They are so informative. I got a nucleic acid biotech paper tomorrow and you're my run to channel 👏🏾👏🏾

[@shomusbiologyofficial]

Thank you so much for appreciating my efforts

[@vishnudasan3711]

what a advance technique,this is not in my syllabus.i had in my syllabus only basic sequencing methods thankyou to make this video to know this advance technique..i love you man you are awesome

[@shomusbiologyofficial]

+Vishnu Dasan well knowing extra is good. All the best. Thank you.

[@danielgladish2502]

Awesome video! I'm going to be using Ion Torrent in my upcoming job and this really helped clarify how the process works! :)

[@shomusbiologyofficial]

You're welcome. Glad to hear that you're getting benefit from my lectures

[@albertkeyan]

You explain the Ion Torrent the best... even better than the corporate video. One question if you can answer, does the chip need to be disposed of every time the sequencing job is done? Or is it reusable?

[@shamsarefin5680]

Stunning video, take a bow.

[@shomusbiologyofficial]

Thank you

[@asmahameed781]

Brilliant! thank you so much shomu for making things easier for us to understand. I am very happy Alhumdulillah.

[@shomusbiologyofficial]

You're welcome

[@carolinecorrea8734]

Thank you for the explanation. It was the first video that I understood everything. Great job. Keep going o/

[@shomusbiologyofficial]

You're welcome. Glad to hear that you're getting benefit from my lectures

[@mohamedhammath4167]

Sir your videos are so helpful to us.. THANK YOU VERY MUCH.... please put a video on ILLUMINA sequencing also ASAP..

[@shomusbiologyofficial]

Glad to hear that you're getting benefit from my lectures

[@atreyeesarkar3035]

if the beads bear multiple of complementory ssdna seq. then there should attach more than one dna in one bead nd they must be different fragments...as a result seq. should be different..tn if we add one single dntp tn in one strand it may not attach but it may be complementory to another fragment attached on the same bead..tn how could we distinguish that from which fragmented dna part this sensing came??

[@Machomanrandyaverage]

with emulsion pcr. Each tiny vessicle of master mix+bead once PCR rxn'd. Then the fully covered beads get loaded post PCR onto the chip and each well senses H+ from the whole bead natural nucleotide incorporation.

[@tomasvalek381]

Everything clear, thank you.

[@shomusbiologyofficial]

You're welcome

[@renanleite4839]

Very good explanation. It was easy to understand because you explained well.

[@deoxene]

Shomu is the best!

[@shomusbiologyofficial]

Thank you so much for appreciating my efforts

[@aliesmailizadeh9610]

Thanks for your excellent teaching. May you share your talk on HIFI technology.

[@hasithbuddhima4803]

you arw the best teacher💞💞💞

[@shomusbiologyofficial]

Thank you so much for appreciating my efforts

[@sibelamisheva3688]

Thank you so much for this great explanation! You are a great teacher! :)

[@adamprabowo4556]

AMAZING!!!!!! Thanks prof!

[@shomusbiologyofficial]

You're welcome

[@MaryamZulfiqar-uk3dv]

It was very well explained

[@shomusbiologyofficial]

Thank you

[@alanlybus4352]

Very good explanation!

[@shomusbiologyofficial]

Thank you

[@thilini79]

Great video. You explain the things very well. Thank you so much....

[@phoenixhouse8075]

Thank you! excellent and helpful tutor!

[@shomusbiologyofficial]

Glad to hear that you are getting benefit from the videos

[@felipevilicich980]

For real I´m going to add you to my graduation speech

[@artigangarekar984]

Thank you sir Helpful video

[@shomusbiologyofficial]

You're welcome

[@longboardordie]

Thanks! This was tremendously helpful!

[@benichauque6888]

Thank you very much for your brilliant explanation. One question: how is the CPU going to distinguish the pH increase caused by incorporation of A from that caused by incorporation of G?

[@berryberri3914]

It does not, there are cycles in which different nucleotides are added to be polymerized (automatic instrument) so the machine just knows which nucleotide it is adding

[@mhtsakosG1]

I am undergraduate student, and i find this video explanation, decent. I have some questions: 1) You draw Emulsion's PCR Beads, 1 ssDNA bind with an adaptor-primer who is bind on Bead surface. Is this possible? I mean, the last stage of Emulsion PCR shouldn't be the extension, so all fragments will be dsDNA? 2) How you wash Emulsion PCR by-products? with streptavidin beads, (biotinylated DNA + Streptavidin-coated magnetic beads),right? 3) These beads you draw, are the Emulsions PCR Beads. Are these glass-beads? 4) According sequencing synthesis, you extend the fragments which are bind with Emulsion PCR Beads? or with Streptavidin-coated magnetic beads? I would apreciate a lot if you answer my questions, thanks a lot for your time #Shomu's Biology

[@nalini007mishra]

These r polyacrl beads

[@georgiakarachle219]

hello Mr. Shomus, I would like to ask: On each bead, are all the Dna fragments that are attached the same or are they form different sites of the genome?

[@faiqaatique534]

well explained! good job

[@arshiafsana472]

Does one well contain one bead or, many beads are there???

[@DomenicaOrtiz-wo8hq]

Thank you so much!

[@shomusbiologyofficial]

You're welcome

[@poojaarchanasahani2873]

you are incredible

[@shomusbiologyofficial]

Thank you

[@the_encephalonaire]

How to perform chip well washing after each dntps? What's the procedure? And what solution is needed to do that?

[@monicagurupriya4105]

Can i get a video on Ilumina sequencing of 16 s rRNA plz..

[@adilsiddiqui123]

very clear explanation!

[@renukamaharjan3483]

Single beads contain multiple DNA fragments attach.. how can we analyse the signal generated and use them for sequencing each fragment

[@alizayahmad656]

How we make fragments single stranded? By adding adaptors?

[@anuradha1132]

what is the exact meaning of sequencing by synthesis??

[@rakeshbelludi7094]

good one bro... try to incorporate some drawbacks of each NGS

[@atreyeesarkar3035]

beads are solid..ok..but when we are loading them to wells are they get attched to wells? if they do so..tn why?? nd if they don't get attached to the surface of the wells then they should wash off too when we are washing out one dntp provided before!

[@YashPatel-si3vq]

Sir can you please make one video for DNase used to treat the cystic fibrosis as therapeutic value

[@shomusbiologyofficial]

Okay

[@ozlemaltay541]

Thanks for your amazing videos I have a question: Are the DNA fragments around one single bead same?

[@amanduggal1]

Hi Ozlem, I am also new to this technolgy, but yes according to my understanding each bead has one type of DNA fragment. Actually one DNA fragment that binds to the bead makes copies of itself and these copies spread across that same bead. I believe this is how it works. Anyone else who is more expert on this technique please correct me if I am wrong.

[@asinjitdas3113]

How would it differentiate between two bases , ????

[@simranpareta9807]

Sir there is no need of primer ?

[@riyaroy7386]

Thank you

[@shomusbiologyofficial]

You're welcome

[@waljat1]

explained really well... can I get a reference for this lecture??? please

[@EasyJay1000]

Thank you!

[@shomusbiologyofficial]

You're welcome

[@shahinmonschizadehtehrany3902]

Very good videos!

[@arsalankhan-cx6bk]

Please please upload illumina sequencing method video

[@isadorah4969]

Should we cut DNA samples in specific ways so we know all fragments are cut at the same place and then somehow separate those known-to-be-equal DNA fragments to run in the sequencing? It seems like the beads might get random sequences of DNA, so the signals would be very messy, right? How is this solved? (I'm a freshman pls don't roast me)

[@sagarrokade49]

But sir how can we get proper DNA sequence from ion sensetive layer🤔

[@siddharthadas86]

i had three questions. Is the PCR step after attachment to the bead? Second do we expect a different pH change for each type of nucleotide? And finally since pH change is going to be a continuous variable, how is it determined what should be a pH change for incorporation of a specific base? Thanks again for the nice explanation.

[@TheWaterstyle]

I think he got the point with the attachment to the beats wrong. I guess each beat has another adaptor and i use pcr to fill the beats. So that there is only one fragment of information on the beat. Another option would be the limited number of adapterpoints on the beat, but i dont know about that one. The pH change is always the same- on H+ per reaction. But i always use just one type of base at the time and change it every 15 seconds. So i start with A, mesure the pH, wash it out and follow with Base G and so on

[@MohorMom]

I was also pondering over the same... I think, the answer to your question might be - "Each microwell containing a template DNA strand to be sequenced is flooded with a "single species of dNTP"... So from ONE well you get response for one type of nucleotide... So each well must be showing the position at which that one kind of dNTP is attaching.

[@sanaali-nn4io]

really really helpfull...! thanku

[@tetsu6strings]

Thank you helpful video!

[@miguelangelcc-wj9132]

Can you please make a video with PacBio seq

[@shomusbiologyofficial]

Okay

[@miguelangelcc-wj9132]

HiFi & long Reads. Please

[@sdfPZXC]

great tutorial

[@GeorgeSandle]

Great video, thanks :)

[@linaki4]

great job!thank you!!!

[@satishvarma4378]

Very good continue boss I like it

[@shomusbiologyofficial]

+Satish Varma thank you. Stay tuned

[@huemanjaimes9291]

muchas gracias¡¡ desde mexico

[@walaaslouma1911]

how we can use it in genotyping

[@rebeccasakell4491]

awesome!!!

[@shomusbiologyofficial]

You're welcome

[@abdullahasad4694]

Plz make a video on Illumina Seq

[@shomusbiologyofficial]

Okay. I will try

[@ahmedhashim5783]

not good.. but how we can know thats was A , B, G or C if from all of them the H+ will reliys??

[@minahilzahid900]

Why don,t you make video on SOLiD sequencing?

[@shomusbiologyofficial]

I will make it soon

[@prakritisundarsamanta4462]

this is best

[@marilenamela9524]

Awesome!

[@hatimmostaf9718]

great thanks alot

[@dbzmoments4262]

Great job ;)

[@floflyberlin7392]

so far so good.. but how a dna sequenz is shown by this. A G T C all create the same change in ph ??

[@Sam-ez3bl]

only one type of dNTP is washed over the wells at the time so the change in pH corresponds to which is present in the well

[@JoanZowlen]

I love it

[@shomusbiologyofficial]

You're welcome

[@vholicek]

Nice vid

[@romennaorem]

great!!

[@lakshmiprasanna9245]

Sir please tell us about SOLiD sequencing

[@shomusbiologyofficial]

+lakshmi prasanna okay. Will do that

[@nopenope7665]

you really are an excellent teacher! i do have a question though - so after the addition of each dNTP, is the cell washed and then the second dNTP added? Also, in terms of of the chip - so how does it work with many samples? does the laboratory need to procure 1 chip for each sample? Thank you again

[@bobdylansmith1]

After you go through each of the four bases, you're not done, are you? Don't you need to keep repeating the four bases to get the entire fragments attached to the bead sequenced?

[@tasbeehazarrin6463]

Sir apki awaz bhut show rahti

[@bernardbagezahayo2717]

chez nous au burundi pour faire la PCR on prélève les echantillons sur les papiers buvards et onrefere au laboratoire nationale de bujumbura et on attend le feed-back