Sanger sequencing method - dideoxy sequencing of DNA

Sanger sequencing method lecture - This lecture explains about the dideoxy chain termination method of DNA sequencing which is also known as Sanger sequencing. The mechanism of Sanger sequencing is explained with this video lecture. In this DNA sequencing approach, modified dideoxy nucleotides are used to cause the chain termination of DNA synthesis which helps to find out the exact DNA sequence. This method of DNA sequencing is named as following -
Sanger sequencing
Dideoxy sequencing
Chain termination method of DNA sequencing.
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Thank you for watching the DNA sequencing lecture on dideoxy chain termination method also known as the Sanger sequencing of DNA.

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[@sarangas8688]

Excellent ! Good explanation But voice is little unclear

[@v.saikiranv.saikiran9752]

Sir please explanation chemicals to indentiy activesite arg, his, lys, his

[@zarlashtkhan3540]

@@shomusbiologyofficial in case of ddgtp how can be a 3 nucleotide sequence formed if both the consective gtps ar ddgtp please answer

[@misha9672]

@@zarlashtkhan3540 it is just because incorporation of ddgtp is a random process. It is not necessary that every complementarity of A is DDG. G can also be complementared. ddntps are provided in low concentration in the mixture than regular nucleotides( A, T,G,C). It means regular nucleotides are also always present. So incorporation of ddgtp is a random probability( sometimes in the beginning complementary to 2nd C, sometimes to the 3rd C, sometimes to the last C). Hope it helps.

[@gerganabonev]

There are all 4 dNTPs added in each reaction and also one of each ddNTP in each well is added because we want to produce fragment with different length. Otherwise the reaction would have stop at the first complementary ddNTP added after the primer. and there will be only 4 length fragments.

[@aboulfazlrad6482]

Exactly and thanks for mention this, otherwise it does not make sense. So I suggest to edit this video.

[@red8527]

Yeah, thanks for clearing that. I was wondering how he got 2 GTP & 3 GTP as well. It should have terminated at 2 GTP position only if 3 dNTPs & one ddNTP were used.

[@sowhatsnew6271]

Thanks .now i am understand

[@thoibathongam5012]

Exactly... Thanx for supplementary this point.

[@manashpratimsarmah4487]

Absolutely right 😊.. I want to write the same thing

[@eugenekim3037]

Shomu knows how to explain DNA sequencing very very effectively. I could not understand the ins and outs of the Sanger DNA sequencing until he went into explaining the method step by step, beginning with the explanation of dideoxy chain termination. I plan to watch all of his videos on DNA sequencing.

[@emilyparks8635]

This is by far the most helpful explanation of Sanger Sequencing; it's a whole walk-through of how/why the process works (rather than just pieces/bits of information from other videos). Thank you very much!

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I am a second yr medical student and i really can't express how helpful this and your other videos on genetics have been to me ! You are a wonderful teacher ! Thanks a lot for teaching the topic so beautifully.

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[@deeprobanerjee7622]

Excellent explanation.. before watching this lecture, I tried to read a review paper on sequencing published in the Genomics journal, watched a lecture by Worcester Polytechnic Institute but you, my friend, are the real deal.. Thanks!!

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Thank you so much ...this is the first time I got the Sanger sequencing ..you put the whole process together in one lecture now its easy to remember the whole story...before I used to learn parts of Sanger sequencing and could not connect different parts..👍

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[@msnadeem837]

Nice presentation. However, as a colleague I would suggest you to add 4 dNTPs plus limiting amount of ddNTP in each reaction tube. Otherwise, you will only get the chain terminated at the first nucleotide such as G, and no next G can be detected....

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Wow, I wish my professor taught this way. Now I understand what I read in the textbook and slides. Thank you

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[@trishabhattacharyya1918]

The best video I found on the internet wrt to Sanger Sequencing. Thank you so much!

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Very well taught up till now I was unable to understand sequencing but after watching this video I cleared my concepts thanku shomu

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this video was so wonderful and these videos open new windows about biology.

[@aadarniya]

4th sem biotech student here. You're awesome. College mein shaam ke 6 baje padhaya thha Sanger Sequencing, kuch nahi samjha. Now I'm quite clear about Sanger Sequencing. Thanks a lot.

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@@shomusbiologyofficial in first test tube why don't we add all the dntps n only three please clear my doubt , appearing for my final MD exams

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It's really help me to understand the actual process of Sanger sequencing...so clearly & easily...

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After reading from notes which is given by sir and also from slide share ... but not clear with concept... n have confusion .. here i am at 2 am watching ur video... n nw 100% clear with basic concept of method.... n clr with all my doubts.... thank u soooo much... 💕

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[@samandissanayake2024]

I always watch your videos. Those are really helpful. And in this case yes as some others have commented, all the four types of dNTP's need to be added otherwise how you read the next A after reading the first A or C after reading the first C.

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[@gvaradan]

Awesome. Tough concept explained very simple. Got confused a bit with why we got 2 and 3 GTP. But after reading posts below I understood the concept.

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Please explain

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I wonder how cud DNA is still synthesized, even after adding ddNTPs in case of C

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[@shafisirbiology760]

@7:00 here not only 3dntps in each reaction centre but all 4dntps and one of each ddntp in each well is added

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point to be noted.

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[@dearbiology]

Great lecture sir but i have a confusion When the unknown chain have two CC so once the chain terminate at the first C theb how it go farward toward the 2nd C

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[@praveenvemuri]

12:44 - Is it possible to differentiate DNA bands in varying length of 1bp on agarose gels?????? The sequence products are analyzed on 6% polyacrylamide gels (very sensitive in differentiating) instead of agarose gels.

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[@meganevans8006]

I can explain the Sanger method now. Thank you!

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Glad to hear that

[@addafusion9539]

Sir for the tube of ddgtp, when there is C in unknown strand then there will be only 2 base in the complementary strand, because there is no normal dgtp present in the second tube when the first ddgtp is added then there will be no chance of adding further nucleotides. Please look into this matter

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[@kavithanair9165]

Kindly clarify , in the tube with ddGTP, the strand formation will end with the first Class it encounters. I.e if the template is ACC, then the resulting strand should be AG. I.e two bases. Then how do we get another strand with three bases.