Sir mujha itni achi samj University mai b nai ai jitni achi apky lecture sy ai ha thankyou so much jazakAllah sir ☺ please asy hi lecture daty rahiya ga ham students k lia please
It's my seminar topic .. thank you so much for helping me to understand the concept .and many people didn't explain about the chemical reagents but did ya .it is very helpful.
To the point , clear, crisp, simple, easy to understand. You might also love to know that there is one particular University in India (whose name will be kept confidential) where one of the Genetics professors is giving notes to her students straight out of ur videos, as it is, without any modifications whatsoever. ;D U shud b proud bruh!
Although i have a question on the G tube and A+G tube. G is the first nucleotide in the example sequence u have used. When a cut is made, it will be made before G, meaning the first cut will produce only the radiolabelled phosphate without any nucleotide. But u still considered this a fragment and counted it as one fragment. Why is that?
I subcribed to ur channel.... I like it... Now I am commenting... Bro... That was truly awesome... U r a great teacher.... Respect from bottom of heart...
Sir, if we don't know the sequence of the DNA then how can we cut with restriction enzyme . As to cut with RE we must know the sequence of a restriction site ?
I have a doubt regarding the 12th nucleotide..if piperidine cuts in C Nd C+T tube it will cut only at 5' of C thn it will become 11 nucleotide ..how it can be the last one my concepts isn't clear there ..thn according to me it lacks one nucleotide
you said only one cut will be made per fragment but then you said 6 or 2 fragments after multiple cuts will be produced can someone clear my confusion please
You said that during degredation every ssDNA will only be cut at a single Position. I am afraid this is wrong. In reality a ssDNa can be cut multiple times. But since only one radioactive phosphor is introduced, you can only see those strands that contain the 5'ending with the radioactive phosphor. So it does not matter if the other end is cut any more since it is "invisible" to us.
10:00 I would like to ask, do we amplify our strand that we will be able to get those number of strands in each tube? As for that to be possible that means we have to have the same DNA strand in each tube. Thank you😊
Thanku sir.. it was very helpful .. but sir i have a question... if dna cut at restriction site and divided into two.. then the smallest one is discarded as you mentioned in the lecture.. so we will not get the sequence for that discarded part? We will get dna sequence for the longer part only..?
Sidra Sanam yes , if the fragment is very small compare to other fragment then we can discard the small fragment and we will not get the sequence of that..... Only sequence larger fragment .
What is the purpose of 3rd step? I am not getting. Suppose after end labelling we denature the DNA we will get end labelled ssDNA.. Then why we are cutting that DNA?
Sardar Zain ... Yes , that is an alternative method , i forgot to mention it but .....in some cases isolation of labelled ssDNA is more difficult by this method . May be that is the reason for including the restriction enzyme digestion step .
This is because denaturing a whole DNA double strand requires more energy, so as to avoid human errors, the DNA has to be prepared into shorter fragments for easier electrophoresing
I don't understand in the step where we are doing dephosphorylation why is hydroxyl group attached to 5 prime end of both the DNA strands that is a bit confusing
Can I do msc in genetic engineering after my Bsc in microbiology. Which goverment University in india allows msc in genetic engineering after bsc Hons in microbiology?
Sir why we Have To elute out the heavier band and work with only the lighter band after the denaturation of DNA double-strand?? Our desired sequence may be in the the lighter band as well isn't it? Please reply sir
Sir in each tube there is one strand and one primer...if once primer cut strand at one position so this fragment separated with primer.then there is no other primer to again attach to template so how next chain start to make 2nd fragmnt???
We run marker DNA in one lane to get the idea about the length of DNA fragments . For more details at first follow videos on agarose gel electrophoresis to clear the concept .
To determine the direction of the strand i.e 5' to 3'...As we know that the phosphate group is only exposed at 5' & not 3' end...So it will help us to pick the exact strand needed for this procedure which is 5' to 3' strand
How we know that dna fragment have 12 nucleotide which we take for experiment or how we know no. Of nucleotide in dna fragment which we take for experiment sir
This is the best video I have seen on this topic so far. So detailing and comprehensive.
i took so many yt lects and did not understand and Alhamdhulilah i found this one its so clear word by word and all reactions wow
Sir mujha itni achi samj University mai b nai ai jitni achi apky lecture sy ai ha thankyou so much jazakAllah sir ☺ please asy hi lecture daty rahiya ga ham students k lia please
It's my seminar topic .. thank you so much for helping me to understand the concept .and many people didn't explain about the chemical reagents but did ya .it is very helpful.
Great sir the best method of teaching very helpful agr sab teacher ap k esab say padae tho kohe subject mushkil nahe oga sab asan say samj ahe gay
Wow! This is simply amazing... My 3 days+ search for clarity has just ended. Well done 👏
Thank you for making me understood this concept..
To the point , clear, crisp, simple, easy to understand.
You might also love to know that there is one particular University in India (whose name will be kept confidential) where one of the Genetics professors is giving notes to her students straight out of ur videos, as it is, without any modifications whatsoever. ;D
U shud b proud bruh!
Although i have a question on the G tube and A+G tube.
G is the first nucleotide in the example sequence u have used. When a cut is made, it will be made before G, meaning the first cut will produce only the radiolabelled phosphate without any nucleotide. But u still considered this a fragment and counted it as one fragment. Why is that?
excellent presentation!
Sir ur lectr is awsom
Thanku so such an amazing video
Gd explanation sir
Thank you very much
what an explanation and points extracted ...thanks a lot sir ...really big thanks ..
Beautifully expalined✌️
Thnk you vry mch sir... It is much more helpful to me than other vdeos.. 😊
It's an awesome video and I don't want to nitpick but, and I am really sorry for this, its "labelling".
Keep up the good work !
Thanks I have understood the technique and continued on like this
excellent sir...... now my concept is clear... n the credit goes to u
I subcribed to ur channel.... I like it... Now I am commenting...
Bro... That was truly awesome...
U r a great teacher.... Respect from bottom of heart...
so nice trick of explain sr thank you so much
Jyoti Sharma .... Most welcome ☺☺
thanks for sporting us
Very good presentation ...
Sir very informative vedio and easy to understand..plz make vedio on automated DNA sequencing
Very good video it helped me a lot thank you wish u luck with scientific projects :)
Matej Forgáč ... Most welcome ☺☺
Thanks sir 🙏
Superb ,thanks for ur clear lecture
Thanku so much sirrr..... Lots of prayers...u nailed it
Thanks sir
Very nice sir
Thankew sir good job..very helpful..keep doing
most welcome
excellent & very helpful
Puja Kumari thanks 😊😊😊
Thank u
Thank you. Very helpful.
Spill Freak 😂😂😂
Nice
If the first base will cut. Then how the dna fragments are formed after it?
I think that's why 32-p i's added,it will produce radioactivity at that region As it carries radioactive phosphate
Multiple copies of the ssdna are added.
Is the same as KDEL sequence?
يسعد دينك اخيراً فهمت
Sir, if we don't know the sequence of the DNA then how can we cut with restriction enzyme . As to cut with RE we must know the sequence of a restriction site ?
I have a doubt regarding the 12th nucleotide..if piperidine cuts in C Nd C+T tube it will cut only at 5' of C thn it will become 11 nucleotide ..how it can be the last one my concepts isn't clear there ..thn according to me it lacks one nucleotide
You stumble a lot Sir but still your way of teaching is very appealing 👍👍.
thkxx u do good work
God bless u MERE DOST
you said only one cut will be made per fragment but then you said 6 or 2 fragments after multiple cuts will be produced can someone clear my confusion please
You said that during degredation every ssDNA will only be cut at a single Position. I am afraid this is wrong. In reality a ssDNa can be cut multiple times. But since only one radioactive phosphor is introduced, you can only see those strands that contain the 5'ending with the radioactive phosphor. So it does not matter if the other end is cut any more since it is "invisible" to us.
u gave the perfect answer of my doubt . thanks
nice explanation
Outstanding boss👌
Why can't we break adenine in the second test tube as we did in the case of guanine?
10:00 I would like to ask, do we amplify our strand that we will be able to get those number of strands in each tube? As for that to be possible that means we have to have the same DNA strand in each tube. Thank you😊
Stephen Lentsa .... We take multiple copies of that DNA strand in each tube .
@@microbioscope18 that is done by PCR?
@@peepdi Yes
🙏Best explaination but sir if there are 12 nucleotide and C is removed by removing agent then largest sequence will be 11 isnt it?
This is amazing my brother . Thank you .
dude... you are the man!
Thank you🙏
Thanku sir.. it was very helpful .. but sir i have a question... if dna cut at restriction site and divided into two.. then the smallest one is discarded as you mentioned in the lecture.. so we will not get the sequence for that discarded part? We will get dna sequence for the longer part only..?
Sidra Sanam yes , if the fragment is very small compare to other fragment then we can discard the small fragment and we will not get the sequence of that..... Only sequence larger fragment .
subrata das thanks alot sir for the reply
Its little confusing when u say its removes the nucleotide. The chemicals doesn't remove the nucleotide but it cuts at after that nucleotide.
Sir can u tell me that in each tube is there many copies of ssDNA is present or for each tube only one strand is present? Can u clear this plz
What is the purpose of 3rd step? I am not getting.
Suppose after end labelling we denature the DNA we will get end labelled ssDNA..
Then why we are cutting that DNA?
Sardar Zain ... Yes , that is an alternative method , i forgot to mention it but .....in some cases isolation of labelled ssDNA is more difficult by this method . May be that is the reason for including the restriction enzyme digestion step .
This is because denaturing a whole DNA double strand requires more energy, so as to avoid human errors, the DNA has to be prepared into shorter fragments for easier electrophoresing
Very good Sir.You have the capability to understand the students . But sir what is most modern technique to sequence DNA?
Next generation Sequencing (NGS)
@@microbioscope18 oxford nanopore is the most advanced or most used is illumina
I don't understand in the step where we are doing dephosphorylation why is hydroxyl group attached to 5 prime end of both the DNA strands that is a bit confusing
Awesome
Why one or two nucleotide neglected during electrophoresis
nice explanatn ...please improve audio
Can I do msc in genetic engineering after my Bsc in microbiology. Which goverment University in india allows msc in genetic engineering after bsc Hons in microbiology?
you are amazing you saved my life thanks new subscriber
Sarah 241Lotus ... Most welcome
Amazing
Sir why we Have To elute out the heavier band and work with only the lighter band after the denaturation of DNA double-strand??
Our desired sequence may be in the the lighter band as well isn't it?
Please reply sir
Sir in each tube there is one strand and one primer...if once primer cut strand at one position so this fragment separated with primer.then there is no other primer to again attach to template so how next chain start to make 2nd fragmnt???
iqra malik ... No , there are multiple copies of the DNA present and there is no primer .... Chemical reagents are there to cut the DNA .
Sir actually my questn is about sanger method...by mistake I coment here...so plz clear it regarding sanger method
subrata das reply must..
iqra malik .... Ok ... In sanger Sequencing there are multiple template DNA and primer is present ... I show only one to explain it easily
subrata das manyy manyyy thankssss
And 13:10 how can we determine the length of the DNA strand on the electrophoresis gel?
We run marker DNA in one lane to get the idea about the length of DNA fragments . For more details at first follow videos on agarose gel electrophoresis to clear the concept .
Thankuuu...its helpfullll
how the same nucleotide will give us different bends in the same tube?
Send the link of Sanger method
Can 1st nucleotide be detected? If not, what is the reason?
Sir Gene mapping video load kijiye na pls
I have one more doubt why are we adding radioactive phosphate in both the DNA strands.. when we are only going to put one strand in each tube
To determine the direction of the strand i.e 5' to 3'...As we know that the phosphate group is only exposed at 5' & not 3' end...So it will help us to pick the exact strand needed for this procedure which is 5' to 3' strand
Also mention your references from where you have taught this to us so that we can know about it
Sir first mey toh Apne Likha hai ki double stranded DNA...usko fir denature kiya.....fir uska sequence kyu banaya....o to replication sey bhi ho jata
Based sequenced method is alse called maxam gilbert sequencing.... It is right or not
where do you get this process??? i mean what is references????
How we know that dna fragment have 12 nucleotide which we take for experiment or how we know no. Of nucleotide in dna fragment which we take for experiment sir
Each band ( in different position) represents one nucleotide ....so total band number denotes total nucleotide number .
@@microbioscope18 thanku sir
amazing bbro, you saved me
hello sir, i have one question
in this sequencing why we use restriction enzyme to cut dna strand if we want to single strand
Because this enzyme cut DNA at specific sit
🤩🤩🤩🤩🤩
Icont usd plzzz
7:09
Jo samjany wala part ta vo tu sahi sy samjaya nhi
chemical degradation not degredation
ayisha moideen .... Yaa ... It's a mistake 😁😁.... Good observation 👍👍
Plzzz replyyy