Maxam gilbert DNA sequencing method

[@mpv6426]

I just realised that I've been passing my entire uni thanks to you but never said a proper thank you so: Thank you! (=

[@Sheikhandbirds]

Same 😂😂😂

[@rosh_serendipity939]

Samee 🥹

[@praveenvemuri]

Its a good try, but there are few things which has to be dealt for better understanding like: 1. what chemicals are used specifically to cleave a certain nucleotide (G-DMS, A-Formic acid, C-Hydrazine+NaCl, T-Hydrazine) together with piperdine to cleave the phosphodiester bond of modified nucleotide 2. How to make single stranded DNA fragments (heat denaturation or using helicase) 3. Dimer formation are in combinations of A+G and C+T If a band in the DNA sequence appears in both the G-reaction and the G+A-reaction lanes, then that the nucleotide is a G. If a band in the DNA sequence appears only in the G+A-reaction lane, then it is an A. The same decision process works for the C-reaction and the C+T-reaction lanes. 4. What quantity of sample DNA is required for sequencing (if not sufficient, have to run PCR or do cloning to amplify) 5. What about the left part of DNA strand after cleavage (they cant be visualized, due to lack of isotope) 6. What percentage of gel is preferred (6% polyacrylamide gels are used, bcz they are too sensitive in differentiating the DNA bands with 1bp size)

[@ProfSardarMNiaz]

Praveen Kumar you are stunning.help me a lot

[@savitapal9943]

hello friends

[@rishabhjain7690]

Praveen Kumar Thanks

[@roshanaatreyapaudel4033]

Thanks

[@yingliweng9759]

thanks

[@postmorton2493]

I'm doing an essay on genomic analysis, and these videos on sequencing are amazing for a proper, scientific understanding of how sequencing has developed over the years. Thank you so much for the clear and helpful video!

[@shomusbiologyofficial]

You're welcome. Glad to hear that you're getting benefit from my lectures. Please subscribe and share

[@ananyayadav9686]

I came across this video , and may i say you are the best youtuber teacher we have got Hands down sir, you are brilliant

[@romi6374]

seriously... The Best Lecturer Out there.. I find it so effective.

[@ejazkhan2952]

no need for short videos....you are doing great job... because it is helpul for begginers,,,if you start to make short videos ...then the beginners will not understand....thanks alot...very helpul.....

[@shomusbiologyofficial]

+Ejaz Khan thank you.

[@sohailvet14]

I am a veterinary student .....GADVASU....number one veterinary university in india......still we dont have any teacher like u sir......thank u very much❤️❣️

[@shomusbiologyofficial]

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

[@drsahivlogs676]

My best BIOLOGY teacher over whole youtube

[@shomusbiologyofficial]

Thank you so much for appreciating my efforts

[@drsahivlogs676]

@@shomusbiologyofficial Love From Pakistan

[@AzhunkWorld]

Interesting and very clear explanation. don't bother about the time because it's very interesting and understandable. Thanks shomu!

[@RosemaryUkamaka-ie7uc]

This is so helpful but I almost got confused at the initial result interpretation but thank God, I later understood it. Thank you Shomu Biology

[@shomusbiologyofficial]

You're welcome

[@reshmirajeev5770]

Thankyou sir.....you re the best teacher...😍

[@KountayDwivedi]

Awesomely explained. This is my first video on Maxam-Gilbert, n you cleared the concept seamlessly. I won't go into technical details, I will just say that concept-wise this lecture is awesome. :-}

[@shomusbiologyofficial]

Thank you so much for appreciating my efforts. Glad to hear that you're getting benefit from my lectures

[@anjiman5890]

@@shomusbiologyofficial,your class is really helpful, sir you take an example of 12 nucleotide, and in autoradiography in the 11th portion the 12th nucleotide come..can you please explain it

[@bamd9]

but what about the first nucleotide ? how to know it?

[@tamannaparvin5797]

Thanks for this video. It's really helpful for me.....This topic is in my syllabus....thanks a lot

[@shomusbiologyofficial]

You're welcome

[@fatimariaz1233]

when you write down on board that method is good for understanding and due to only this difference between you and other you tubers i was watching your videos

[@CringeDealer69]

Really really helpful... Understood the topic thoroughly... ☺️

[@shomusbiologyofficial]

+mayur tupe thank you. Glad you liked my lectures

[@CringeDealer69]

Shomu's Biology My pleasure... ☺️

[@alwinaanam1231]

indeed its helpful...........complicated topic bt after watching twice /thrice i can understand for sure..

[@shomusbiologyofficial]

+Alwina Anam thank you very much

[@nothingggg656]

Thankyou for the wonderful explanation sir❤❤

[@shomusbiologyofficial]

You're welcome

[@nohabassiouny6849]

you have a simple way in teaching and presenting the idea

[@shomusbiologyofficial]

Thank you. Glad you liked my lectures

[@anjalisharma7394]

It’s very clear nd helpful .thank you for this🙏🏻

[@shomusbiologyofficial]

You're welcome

[@nemapusai]

Very nice sir ! no words to tell and your classes helped a lot. thank you soo much sir

[@shomusbiologyofficial]

Thank you so much for appreciating my efforts

[@princecudjoe5518]

that was some top explanation man. thamks!

[@shomusbiologyofficial]

You're welcome

[@jaydeepdumadiya2464]

Beautiful sir... God bless you.. 🙏🙏🙏🙏🙏🙏🙏

[@shahbaloch4814]

Great sir very easy and understanding english best method of teaching 🇵🇰

[@shomusbiologyofficial]

Thank you so much for appreciating my efforts. Glad to hear that you're getting benefit from my lectures

[@arshiafsana472]

And what about first base Adenine...gel doesn't have this info.. so whats the reason sir..plzz...tell us

[@rafaelkyranas3131]

Dude you are the BEST! KEEP IT UP

[@ankitarout2688]

It's really helpful.. Thank you so much

[@shomusbiologyofficial]

You're welcome

[@Serwada237]

Better than my class work ❤🎉

[@shomusbiologyofficial]

You're welcome

[@Serwada237]

Iam so happy that you replied to me I have used your site to undertand concepts for my molecular biology masters degree. At Makerere university Uganda… thank you we appreciate your efforts to teach the world science

[@marwahussien718]

thank's very much now I depend on you very much to understand any thing .I wish I Can reply the favour ❤❤ and please don't stop many students depend on you . Marwa from egypt

[@rumibhadoria4009]

Sir your channel is v helpful . I get everything here

[@adarsh7221]

How can you tell the first nucleotide is A(adenine) from electrophoresis? Please answer

[@asmasbiology3]

yes plz

[@udehedith7326]

The video was quite helpful and made understanding easier. Thank you very much

[@shomusbiologyofficial]

You're welcome

[@ashwathy196]

U are a super teacher 😍🙏

[@shomusbiologyofficial]

Thank you.

[@techforever1970]

Thank you so much! You helped me alot. I owe you.

[@shomusbiologyofficial]

Thank you. Glad you liked my lectures

[@khalidmehsud54]

Hain?

[@ayanasudheer2934]

How did you know that the very first base in the gel is Adenine?? Someone pls answer

[@verdaan_]

I think whole biotechnology students are graduating seeing your videos ❤

[@shomusbiologyofficial]

Thank you so much for appreciating my efforts

[@mahiratariq7864]

Yeah🥰

[@Akshita23436]

Absolutely right

[@hadogori4613]

I love your explanation♥️ Thank you so much ♥️

[@umairameer6728]

How can you identify A at Terminal 5 prime end. As it shows the band in T+C.

[@malleshdurgam568]

Thank you very much sir... 🙏

[@shomusbiologyofficial]

You're welcome

[@monicasingh4156]

It's really helpful...

[@shomusbiologyofficial]

Thank you. Glad you liked my lectures

[@tusharsujgure7379]

Thank you so much sir. Your teaching method is very easy and very helpful for understanding. Sir please make a video lecture on Evolutionary Game Theory.

[@shomusbiologyofficial]

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

[@tusharsujgure7379]

@@shomusbiologyofficial sir please make video on Evolutionary Game Theory

[@kundandutta710]

Sir why we Have To elute out the heavier band and work with only the lighter band after the denaturation of DNA double-strand?? Please reply sir

[@shubhamphatangare6062]

Maxan and Gilbert sequencing explain very easily.. Suman sir please make video lecture on Game Theory..

[@shomusbiologyofficial]

Thank you so much for appreciating my efforts. Glad to hear that you're getting benefit from my lectures

[@shubhamphatangare6062]

@@shomusbiologyofficial sir please make video lecture on Game Theory

[@darshan00185]

Sir it was very helpful but how A is the 1st nucleotide is not I think explained

[@aakaankshavangala1826]

1. Not Chemical Synthesis. It is Chemical Cleavage (two completely opposite meanings) 2. First nucleotide determination without which DNA sequencing is incomplete. From my understanding, you are supposed to initially cut/cleave the DNA sample with a restriction enzyme. From this you get to know where the DNA is getting cleaved (as you know the restriction sequence of the enzyme). This is then followed by the steps explained in the video.

[@sukhmanpreetkaur7951]

can u plz refer me any book for this topic

[@bhagyajitdas1498]

Thank you sir for explaining it beautifully!

[@shomusbiologyofficial]

You're welcome

[@md.shaminurrahman8758]

You are the best💜💜💜

[@sushantikasubbiah1931]

During chemical termination is the single strand added newly to each test tube everytime?

[@prempratyushmangraj1050]

Sir.. How can we know about the first nucleotide?

[@Komall30]

diagram is from which book? a good informative video!!!

[@top-uptv7010]

Please what is the name of the referenced book, how do we get it

[@taniagoyal3523]

Won't their be be combinations for cleaving out T in 3rd tube Like 1 and 2 T And another one 1 and 3 , 1 and 4, 2and 3, 2 and 4?

[@bhargavibitra4593]

Thank you soo much professor 🙂

[@shomusbiologyofficial]

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

[@kavitachoudhary6168]

Sir I have a question As we can see that while chemical degradation sequence is cleaved at the site just before to that perticular residue so what about A which is radiolabled as a first residue. How we will know that A is present at first becs when it get cleaved we will get only phosphate without sequence?

[@chandrat1221]

Thanks! what is the sequencing QiaSeq target amplicon method different from Sanger sequencing method ? which is superior?

[@jerry-pc6fl]

but how can we get the first base by using this method

[@anjalisetia8672]

why the first base (A) is not identified..?

[@kushaniakshala3758]

Great...... Thank you

[@shomusbiologyofficial]

You're welcome

[@duraiswamy8027]

How same DMS cut each nucleotide at different tubes sir ???

[@manishmehra9748]

sir why are we unable to find the first nucleotide ?

[@swatipatil9156]

Bamh1 recognise 6 base pair of DNA in palindromic sequence if the first part sequence is given below what is rest of it? 5 C A A3

[@duraiswamy8027]

At 1 guanine reaction ,u told that DMS will cleave the guanine ,so no stretch of DNA will form . But how it produce 3 types of strand at guanine reaction ,because DMS will break sequence ,so we will get same type stretch DNA only sir ,???

[@magnfiyerlmoro3301]

what do you mean by homogenous dna please thanks

[@nzolelejuma8167]

Sorry sir can you tell me a little beat in interpretation of result sequence if you have C and G of the same size what base will you labble first

[@shreyarastogi4925]

Can chemical sequencing be used for RNA sequence?

[@RafaelMazenett]

Awesome. It helped me a looooot. THANK YOU

[@sheikhabrar9932]

praveen Kumar.....thanks for adding more information

[@nishantsen6820]

Cysteine.....😅 repeated twice lol But nice explanation as always your videos are helpful keep it up👍

[@nabeelkhan-yh9rx]

sir carry on your vedios are very helpful

[@muskaankhalifa5059]

Sir, which book to refer for this?

[@jaivardhantiwari6172]

amazing video

[@shomusbiologyofficial]

Thank you

[@swetapanwar6993]

Sorry sir...i m not able to getting this in last...how we do count ...if we don't know the sequence of complementary then how we count the Nucleotide bases....i saw this video repeatedly but i didn't get it after electrophoresis ..... please tell

[@tunerd1921]

What was the book called?

[@dr.jyotisaxena6966]

Like many others I also want to know whether 1st base can be detected correctly from this method or not.......if not why

[@mssazeb]

i have a question.. why we select G and C bases separately.. i mean we took purines and pyrimidines in two test tubes n G and C in rest of the two so why not A and T??? and we treated T+C tube with the chemical to cleave T and upto some extent C as well.. so why that chemical isn't working for T?? n why the result is same for C and T+C???? reply

[@Obd9900]

doing a great job man (Y)

[@samjhanastha6647]

extremely helpful!!!

[@shomusbiologyofficial]

+samjhana stha thank you. Glad you liked my lectures

[@SedighehMonavari]

great explanation. thanks :)

[@riichobamin7612]

In the results interpretation part, you appear to be removing even terminal nucleotides (like you removed the C from the T+C tube). Didn't you say that the terminal nucleotides cannot be removed ? Also in what manner are the nucleotides being fragmented ? There should be atleast 4 different types of fragments in the G tube for example if we remove the G from the middle.

[@simranpareta9807]

Sir can you recommend books related to life science

[@shomusbiologyofficial]

Campbell Biology

[@shummu1458]

plzzz sir explain this in black board . its a humble request from my side because i saw this explanation in black board on many other u tube channel's. but for me sir only u have abality to explaining in black board with ease. otherwise this presentation is also very good sir👍👍

[@pratitidas]

Can u give the link of the biochemistry book

[@marzooqsalahuddin5935]

Sir can't a cleaved dna be further cleaved ?

[@supriyohalder7889]

thank you bro i got help from this video.. keep it up

[@shomusbiologyofficial]

Thank you

[@maitypartha]

How can find tha 3'A seq then ?

[@wassimsouissi8330]

Does each reaction require many single-strand DNAs ? ( cause there is too many fragments )

[@travell_withheart]

I have understood all but why shoud we underline some particular sequences

[@divyangdamor7380]

thank you... great explanation...

[@subhamsaha2235]

waah sir...lovely...very easy

[@aboutstudies123]

Thank you very much Sir. It helped me a lot

[@savaninagarkar7395]

What is homogenous ssDNA?

[@devsbiology4808]

Sir where is the link????

[@shuvashispaul7330]

Thank you

[@shomusbiologyofficial]

You're welcome

[@mahiratariq7864]

Thanks♥

[@shomusbiologyofficial]

You're welcome

[@sabinahaque664]

Thnk u for helpful explain

[@shomusbiologyofficial]

You're welcome

[@Trunks9Thousand]

How do you get enough fragments? Standard PCR?

[@vinceb8041]

PCR of your sequence gets you lots of fragments, alternatively you could clone it into a vector, insert it into E.coli and let the cells amplify your plasmid. Once you have your DNA, [add 3M NaOH] or [heat to about 90° and put on ice directly afterwards] both methods will yield ssDNA. :)

[@DiegoDiego1989]

I love this guy

[@AaRTiSharma-qs7em]

simply awesome!! got it :)

[@afnansalah4031]

you are AMAZING

[@supritadash2535]

Thank u so much.. really helpful

[@shomusbiologyofficial]

+Suprita Dash Glad it helped