Sanger sequencing method - dideoxy sequencing of DNA
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- Опубликовано: 27 авг 2024
- 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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Excellent !
Good explanation
But voice is little unclear
Sir please explanation chemicals to indentiy activesite arg, his, lys, his
@@shomusbiologyofficial in case of ddgtp how can be a 3 nucleotide sequence formed if both the consective gtps ar ddgtp please answer
@@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.
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.
Exactly and thanks for mention this, otherwise it does not make sense. So I suggest to edit this video.
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.
Thanks .now i am understand
Exactly... Thanx for supplementary this point.
Absolutely right 😊.. I want to write the same thing
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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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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.
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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....
EXACTLY .I WAS CONFUSED THERE!!
Doing a masters in cancer and I can't tell you how much you have helped simplify topics.
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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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Thankyou so much , your videos have been helping me since my bachelors and now my masters as well
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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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Hi sir...Without you and your channel i couldn't get pass in the subject..For each and every semester I'm learning from u..ur trying hard to make us understand so easily..Thanks a lot for your effort...I became your fan since 2017....If u reply i ll be happy...Thank u sir...
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It's really help me to understand the actual process of Sanger sequencing...so clearly & easily...
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Your videos helped me a lot during my PhD journey. May God bless you. Thank you
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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yes he is😊
You explained a difficult concept in a beautiful, simple way. Thank you!
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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
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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Thank you very much, i study biomedical lab.teknologi, and i startet in 2017, and your videos have been very helpful for me, i appreciate it ,and thank you very mig wee love you
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You are blessing, i wish i get teacher like you in university, u clear every single confusing without even asking!
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I couldn't understand it when my professor was explaining it. But you've just explain AMAZING!!! You are the best!!!!
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@7:00 here not only 3dntps in each reaction centre but all 4dntps and one of each ddntp in each well is added
point to be noted.
This guy is THE BEST!!! Thank you soooo much.
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you are the best teacher of bio . Thank you sir for best video.
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Thankyou sir,
Very effective class💙
May Allah bless you with more and more knowledge
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This was great sir! Very well explained. Thank you so much for this precise explanation.
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This is the best lecture. I understand every step very well when you explain . Thank you very much for making it easy and understandable!
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exactly what my teacher trying to teach us. Now it is clear to me.
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You are doing very good work keep doing this work it helps a lot people
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amazing! I have biology paper tomorrow. Your lecture is so helpful
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I can explain the Sanger method now. Thank you!
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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.
Thank you
Please explain
Very helpful and best method of teaching Sir ap bohoth he Acha samjathay he hor padathay he
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Sir you are the best. I have no. Words to explain. You make it understand so simple and explains the best.❤️😇
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this video was so wonderful and these videos open new windows about biology.
Precise and straight to the point, well understood
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I just couldn't thank you enough. Thank you Sir.
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thank you sir for explaining this in simple and understandable manner
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I enjoy your way of talking and helps me to get ur every point.
From pakistan.
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very useful.most of the lectures well describes the steps untill autoradiography,for an unexpert person from that step hurdles starts.this videos thoroughly clarified such a gassy knowledge.thank you so much
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i'm Meherima from bangladesh , a student of Genetic engineering....just love your way of all videos ,these are really so helpful Shomu, God bless you....thank u so much for all these contributions....
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Explained in very easy and grasping terms. Thank youu!
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Thank you sir.... I can't understand this experiment in my class.... I'm struggling to understanding this but not able...but after watching this video I understand this clearly
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You are really saving my ass in my clinical imaging course with these awesome lectures man. Thanks for the good and easy to understand explanations! Liked and subscribed.
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OMG! Just easily explained! You just gained another subscriber!
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I have watched ur many videos bt dis time u really helped me before one day of examination...thanks from nepal
Amazing , helped simplify the concept and really helped me understand!!
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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
The best video I found on the internet wrt to Sanger Sequencing. Thank you so much!
I learn this easily from you....
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Sir u r a genius . It was Soo difficult for me and u made it Soo easy
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God of biology. You are great
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Thank u sooo much sir for ur vedios....i m fully dependent on u for my 12th board....no one can teach biotechnology better than u.....
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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very well explained sir, thank you.
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Thank u so much Sir you explain concept so well
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Thanks @Shomu's Biology
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When the chain get terminated at 2nd position ( C is present)then why will it again proceed to terminate at 3rd position ( again C is present)?
Hey bro ur question is valid.
Shomu has taught in a little wrong way in 1st test tube there should be both ddATP and dATP thn by probability either of them would attach
@@amishbutle4887 yes bro 👍
Same question I wanted to type but after seeing your answer I understood 👍😊
@@amishbutle4887 thanks bro..I also have same question..
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Perfect explanation... I clearly understand it. Thank you so much sir. 🙏
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Far better than any one of my teachers 🙂🔥
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Wow...you are such a great great teacher...! Kudos to you!!! Thank you very much!!!
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Wow... I understood everything thank you so much.
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Thanks a lot, sir. These videos really help us get through college. 🥺😄
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in all the tubes all dNTP's are added, otherwise how would the chains be of different length. For eg in 1st tube according to your method, the chain should terminate at the 1st A attachment only. Thus all chains would be of same length
same remark !
Exactly
Each tube needs one ddnucleotide sequence for termination. Different sequence undergoes different length if they have fluctuating sequence. This is only applied for short sequences, and worth of a Nobel price😊
Each tube have all narmal dntps but one specific ddntps.
abhishek yadav the quantity of di deoxyNTP are lesser as compare to the dNTP.
Hence there are chances of getting dd on the right place as well as d.
During electrophoresis it can give larger bands if the dNTPs are added everytime but ddNTP can also bind
U made my assignment easy tq sooo much sir
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It's a great explanation, but as other comments have mentioned, you are a little ambiguous when explaining how the sequence can continue once the ddNucleotide is added. It is necessary to emphazise that each of the four mixtures contains ALL FOUR dNucleotide PLUS one of the four ddNucleotide.
Otherwise, thanks for the clear explanation.
You Clarified further, Kudos!
thaks .. u cleared my confusion .. about d/ff sizes of sequence.....
thanks！
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Great explaintion You are a great teacher
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Instead of using only ddNTP we also use the same nucleotide otherwise a reaction for ddGTP will stop at 2nd step only and it will not move further to 3rd or 7th step without a free OH group.
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A lot of Thank sir i have no idea for dna sequencing but now i feel confident bcoz of you sir......
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All dNTPs along with 1 particular ddDNTP is added in each tube. As a result of chain termination DNA fragments of different lengths are formed.
Otherwise, if we are adding only ddNTP along with 3 dNTP, termination will occur only in the first one.
How different length fragments are formed by adding same ddNTP in particular reaction mixture the first fragment is terminated due to particular ddNTP attachment,why don't the same ddNTP attached at the same position again, why it's not happening? Will u plss explain to me