Southern blotting
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- čas přidán 7. 06. 2016
- 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 am also biotechnology student in msc..
Exactly 💯
I also
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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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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😑
Very comprehensive.
I am a doctor and you have very well explained about all blotting methods.Thank you
Glad to hear that you're getting benefit from my lectures
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!
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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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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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.
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And thank you for all of you effort too.
Student from Thailand.
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!
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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Excelent explanation! Very easy to follow and very clear. Thank you so much!
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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!
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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Southern blotting was a foreign concept to me before I watched this video. Thank you, Shamu....as usual!
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From Sri Lanka 🇱🇰
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Ameen
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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 ???
again after searching so many sites ,I have reached to the best site .....best explanation
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Thank you, this helped me a lot, and it was explained very well.
I am a medical student and you explained it so well. Thank you
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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 CZcams. 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.
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@@shomusbiologyofficial Welcome sir
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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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Glad to help you out. All the best 👍
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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.
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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Very nice explanation.
Very much easy to understand.
Thanks
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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 ☺️
God, you are a genius ! Thank you so much! I really love your pasion to explain.. I really appreciate your work !
Thank you so much for the detailed teaching methodology of yours....I am much benifited...
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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
Thank you so much for this very helpful video.
You made this much easier to understand, God bless you sir.
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awesome explanation which is not in our practical classes.. thank you very much..I love you man..
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Your lecture are easy to understand and you are amazing 👏
Thank you
Beautiful video you made about southern blotting
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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Thank you for the excellent explanation. :)
Couldn't go back without commenting,, Amazing teaching!!!!👌
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Your video is so much helpful for better understanding
I clearly understood southern blotting from Ur video thank you 😊 so much
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