Southern blotting
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- Опубліковано 20 сер 2024
- Southern blotting - This lecture explains about the southern blotting procedure and technique to identify specific DNA fragment from a mixture of DNA molecules separated by the Agarose gel electrophoresis. Southern blotting step by step explanation is provided to understand the procedure of the DNA separation and identification. Southern blotting application is also explained in this video.
Southern blotting consists of three different stages -
1. First is the isolation of DNA from the target cell.
2. Then the treatment of restriction digestion of the DNA to generate DNA fragments
3. Then the separation of the DNA fragments using Agarose gel electrophoresis
4. Transfer of the Agarose gel DNA bands to a nitrocellulose Membrane filter. This filter paper after the DNA transfer is known as southern blot.
5. Then the use of complementary DNA probe to bind with the target DNA that will indicate the presence of target sequence by radioactivity or colorize trick expression. Southern blotting was invented by EM Southern on 1975 at it was named after him to identify DNA using blotting technique.
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Thank you for watching southern blotting lecture.
Sometimes I think why don't we get professor like you in university?
U r like a....... Complete this line friends
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Blessing ❤
I'm a student of biotechnology. I found that ur 100time's better than my prof. Thank you sir. May god bless you.
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Exactly 💯
I also
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SAYING "THANK U" WILL NEVER ACTUALLY THANK YOU SIR.
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You are a machine my friend. I've watched a lot of videos about SB but a clear explanation of the principle of blotting was nowhere to find until I found your video. Thank you for helping me and you have my respect for what you do here. Sharing and explaining the information is an honorable thing to do. I am glad that there are such people as you.
I LOVE your accent. I can easily understand because you point out all the important parts with accent. And also you know what kind of questions listeners would have and you answered all of them. It is like taking a private lesson. Thank you so much! From Korea :)
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His intonation is appreciable!
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I am a doctor and you have very well explained about all blotting methods.Thank you
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Same here. He is so on point
I think the lab technicians would know more about blotting than doctors? Because it's literally what they do everyday!
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Thank you sooo much sir for your efforts .... i must say that you are the best biology teacher i ever have
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Abhinav goyal sir ka naam suna ha ?
I have an exam on Thursday on this and I never understood this process until I listened to you explain it. Thank you so so much for your help❤️
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This is the first ever lecture that I cried out of satisfaction ! As I have quite being long struggling for this topic .Thank you so much sir.😊😊😊
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Thank you for making this video. You've explained this way better than my prof!
+Laura Prince thank you very much.
My teacher finished in 5 min and i was not able to understand😑
Sir you are a savior of my life specially before exam . You teach every portion in a very simplified and lucid way . Always keep us helping like this. Thank you sir.
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Whenever i need to clear my doubts i type the topic name in youtube followed by shomu’s biology because literally nobody clears my concepts like he does , he is a blessing sent to us by gods 😭😭
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again after searching so many sites ,I have reached to the best site .....best explanation
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Great video! I have Southern Blotting as the subject for my biology presentation, and this is the only thing that helped me understand what Southern Blotting is. Thank you!
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New subscriber here....I always left out the genetics topics without studing in detail in school, even in puc and degree....but now when I started knowing about genetics in master's degree, your videos helped alot for me to understand the basics and made me to learn and know more about molecular biology and genetics...
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No professors explained what southern blotting really is. So you are the best professor for me. Thank you 😊
Good luck sir,
From Sri Lanka 🇱🇰
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You are a life savior sir! I tried to understand this multiple times for 1and half month couldn't get but now I have got it clearly. U just saved me
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i have been watching your vids since many years now.. and you have really improved in ur explanations... ur older vids are nice as well.. but the newer ones are even better.. i prefer this format over the older s.blot video!
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Thank you very very much. There is no such explanation in the youtube even in 2022. May God bless you.
Thank you
I had a chemistry exam today at uni and now i am studying for a molec genetics exam for tmrw and i just wanted to say that god may bless him. 7 yeqrs later and he has helped countless of students.
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I am a medical student and you explained it so well. Thank you
You're welcome
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Ameen
Tnx for these excellent way to explain topics that makes them interesting.
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it took more time to read d same thing from book but just some minutes r required to understand d same thing in more clear way from your lectures...the way u taught d thing and make us learnt is really awesome.
Thanks bro for simplifying such techniques so efficiently..It is far better than those complex contents given in the books....
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I know I'm late but I have an exam tomorrow (Btech Biotechnology) and this is simply the best video on Southern Blotting on UA-cam. Not only the content but the enthusiasm with which you explain is so good to see that I understood everything and am left with a smile on my face. Hats off to you sir!
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thank you sir... your video has actually cleared all my concepts... this is the first time when i actually understood the exact meaning of probe n need of using nitro cellulose filter..... thanks a lot sir... god bless u.
+Meenu Garg thank you. Glad it helped.
u r that friend who helps us during xams by explaining everything for a person like me who is a visual thinker. thanx a ton buddy
+Ann Martin thank you. Glad you liked my lectures
The video made me understand southern blotting very easily. Off to watch northern blotting! Looking forward to more of your content!
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Amazing teacher.....
Thank you soo much sir...and respect u a lot.....
+Amrita Rathore thank you. Glad you liked my lectures
thank you very much. i have my exams the day after tomorrow and this really helped.
Mr Shomu I had a presentation for university about Southern blot , I didn't get it at all before but ur video literally was satisfying and amazing, thank u sooo much man , I did a rly nice presentation today all based on your video with the same way of explication ❤
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I have check severals youtube video looking for some tutoral for my master in microbiology, as a physician i find this concepts very difficult to understand, I am a teacher myself i can notice this is pretty good explaining material you just dont explain the technique itself you relate everything in a way that is easy to learn and you also explain for what is gonna get usefull. Kudos to you. Wish me an approval in my exam
Glad to help you out. All the best 👍
excellant video... even books also dont explain this well. thanks...
+indranil bhattacharjee thank you very much. Glad you liked this lecture.
Thank u sir for getting concepts clear within minutes..
This video was so helpful! It was the best explanation on Southern blotting I have ever seen. Thank you so much!
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Today i learnt southren blotting by this video... 😁😍
Yesterday i learnt Western blotting. Hats off for uploading these type of informative lectures.
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Excelent explanation! Very easy to follow and very clear. Thank you so much!
I think there is no other person who explains like you in a simpler way Even though it is complicated.....
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Thank you so much sir❤ your way of teaching is very understandable..ALLAH pak bless you🤲
Thank you
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+Amisha Patel glad to hear that
Hello Shomu Bhattacharjee !
I'm a 2nd year student who had to deliver seminar about the same topic .The whole concept of SBT has been told by you in a very easy and understandable manner , you video has been very beneficial for me , Thanking you for the same .
Glad it helped
Very nice explanation.
Very much easy to understand.
Thanks
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an amazing video!!!!! thank you very much!!!!
+Joanna Przybyszewska glad you liked my lectures
Thank you soo much sir...I love Ur passion about teaching..
Glad to hear that
Southern blotting was a foreign concept to me before I watched this video. Thank you, Shamu....as usual!
you're the number 1 reason i'll manage to get a degree. i'm your biggest fan!
hope someday i'll learn to be such a great teacher as you are
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@@shomusbiologyofficial
Hmm
Wow what a lecture..♥️
Thank you
Denaturation of fragmented dsDNA is done before transferring to NItro cellulose membrane. Otherwise how this DNA will attach to the membrane.
U r right it should have done before tranfering into nitogenous paper
Hey can you tell me why those ss dna dont bind to each other again before binding to probe ???
bro u r a thousand time better than professors in my university
god bless u
Thank you so much
Thank you Dr. Shomus you are amazing.
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please keep doing these stuffs
+shivam jar thank you. Glad you liked my lectures
Great man
Sir you are the best. I have followed you for my icar jrf exam. Now I am following you to complete my masters in plant pathology ☺☺☺
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You have made a great explanation sir.Your technique of teaching is very effective that clears many concepts. Thank you so much.
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Why do you need alkaline buffer? What would happen if I used acidic buffer, water or Histidine? Why does it HAVE to be alkaline buffer for the imprint to work?
Thank you ^.^ I understand now
dna is negatively charged(phosphate group). alkaline buffer is more basic (ready to share an H+) and acidic buffer is ready to take an H+. in this reaction dna is the acid and is reacting with the basic buffer... this is my thinking..and reference from my orgo chem class... i think the phosphate group which consists of P(-)H3, where the phosphate atom is negative, would give away other H if combined with something more acidic or some compound wanting a H more. Causing the dna to become more reactive.
Acids contain excess H+ ions, which bind quickly to the phosphate (PO4)- backbone and cancel out the negative charge. Without charge, DNA can't migrate because it won't be attracted to the (+) electrode. The alkaline buffer does the opposite, neutralizing any H+ (or Ca2+, Mg2+, or other + ion) by pulling them off the DNA, into solution. Alkaline means excess OH-, so by dissolving any (+) ions, it makes sure that the DNA will be (-) and be attracted to the positive electrode (allowing migration and development of bands).
Sir I am have a doubt.
How do we know that the DNA of the required sequence is 100% present?
Cause we have prepared complementary probe of that sequence right? How do we know that's sequence is present? :/
Also we can do sequencing to know whether it's there or not? I just don't understand.
And also sir can you please explain regarding restriction mapping and restriction digestion. Cause I've watch all of those videos but you haven't mentioned about the partial digest double digest and all. And no one can explain better than you its a requesttttttttttt. And i am in th ir d year biotechnology can you suggest reference books for RECOMBINANT DNA TECHNOLOGY.
actually as far as i know we do sB in order to confirm the gene of interest.
we have gene banks where we have sequenced the genome
and we make the probes against the segments whose secquuences we have already done
and because of hybridization technique ie probes we can diagnose
I hope u have got it
this fanda is diagnosis ☺️
Your lecture are easy to understand and you are amazing 👏
Thank you
Thank you sir soo much due to this lock down I was not able to understand blotting in online lectures from my college but now you have completely made me understand.
You're welcome. Glad to hear that you're getting benefit from my lectures
But what if the restriction endonuclease (DNase I ) cuts in between the target DNA , Then how our probe will bind to target DNA?
megha sharma the restriction endonuclease cuts in specific places only. If you know the sequence of the target dna then you can control which endonuclease to use to cut in what location.
creo que la enzima de restricción solo corta en ciertas partes.
Don't we use nylon membrane in Southern Blotting and nitrocellulose in Western Blotting? Im confused..
Aishwarya D my same initial thought . can anybody explain ? I'm confused
Aishwarya D either nitrocellulose or nylon membrane can be used in both the technique .
both can be used
This the mixture of electrophoresis and pcr😅😅😅 at the middle the blotting done...thank u so much sir... great explanation of each and every steps❤❤❤
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Thank you so much sir.. your very talented and u can teach very clearly and simply.. Better than some prof.s Well done good job.. Again THANK YOU SOO MUCH..!!!
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Your like an gurdian angel sir ...u explain so well .....i watched approximately all of ur videos it's soo easy to understand 💕💕 thank you soo much sir.....✨✨
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Thank you so much for this lecture sir.. Love from Tamil Nadu 😇😇🙏🙏
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Awesome video :) Keep up the great work! Im a doctor and I love teaching too, and your videos show me how to be a better teacher. Cheers!
Finished the video, thanks so much again. Can't stop complimenting you. Such a job has been made... I hope you're continue progressing, evolving.
you are entire indian biotech student's best teacher....
Thank you so much, you are one of the best Biologist
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U invent eastern blotting I will be very happy 😁😁😁👍👍👍👍👍👍👍 God bless you always
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@@shomusbiologyofficial yeh koi system lagaya hai hr bt pe yeh hi reply krty ho
Super sir in this lockdown I started seeing your vedios and I could understand clearly 🙏🏻🙏🏻🙏🏻
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You have so much knowledge and the best thing is you know how to share it as well, keep up the good great.. you're really amazing 💎💎
I'm from Bangladesh.🇧🇩
Sir, I watch your most of cls. U r really amazing
That video was awesome. Gave full understanding. Keep up good work brother
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Wow!! now I finally understand what is Southern blotting. All thanks to you. Ab finally sb smjh agya
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Thank you for making this video. I stumbled on it today and I am definitely subscribing.
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i have no words to say thanks to u ...thank u soooooooo much sir for this effort ..Allah ap ko izat r kamyabia dy
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Your videos are really amazing and life saving for exam preparation.
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thank you so much for your best presentation and clarification
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স্যার আপনি অনেক ভালো করে conceptual বোঝান এতে আমার অনেক প্রবলেম গুলো clear হয়ে যায়।। thank u sir
Dhonyobaad
May God bless you in abundance.... Your videos are really helpful... Thanksss a ton sir.
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Sir once more thank u v much for being such a great help in my career
You're welcome
Your lectures are so much concept clearing sir
Two doubts i have from this lecture
1. How to place the gel on the buffer as it is fragile??
2. How to confirm that all the DNA fragments from gel had migrated to the nitrocellulose paper??
Thank you for clearing concepts of me and many lakhs of students