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- เผยแพร่เมื่อ 17 พ.ค. 2024
- This video explores the basics of Sanger sequencing and the fascinating history behind this groundbreaking technology. It explains the basics of DNA sequencing, including the components necessary for the reaction and the steps involved. Topics also include ddNTPs, cycle sequencing, fluorescent dyes and capillary electrophoresis.
We'll also compare Sanger sequencing to other sequencing technologies, such as NGS, to give you a better understanding of how it all works. Throughout the video, we'll take a chronological approach to explore the history of Sanger sequencing, starting with its development in the late 1970s and following its evolution. By the end of the video, you'll deeply understand Sanger sequencing and its role in sequencing the human genome.
So grab a cup of coffee and journey through the fascinating world of Sanger sequencing! Whether you're a student, researcher, or just curious about the science behind DNA sequencing, this video is for you!
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👉 You may also like: "Next Generation Sequencing - A Step-By-Step Guide to DNA Sequencing." • Next Generation Sequen...
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CHAPTERS:
00:00 The 1977 Invention of Sanger Sequencing
00:23 The Basics of How DNA Is Copied
00:36 What are dNTPs & ddNTPs anyway?
01:19 How Does a ddNTP Stop DNA Copying?
01:54 The DNA Naming Convention 5' to 3'
02:43 Which Bases Go Together in Base Pairing?
03:05 Steps of the First Sanger Sequencing Method
06:11 The First DNA Sequencing Instrument, the AB370A.
07:20 The Launch of the Human Genome Project
07:46 How Does Cycle Sequencing Improve Things?
08:59 Automated DNA Separation by Capillary Electrophoresis
09:36 ABI PRISM 310, Modern Sanger Sequencing
10:49 Fluorescent ddNTPs and the Invention of BigDyes
11:33 The Instrument That Sequenced the Genome
13:09 Why Use Sanger Sequencing When There's NGS?
#ClevaLab #sequencing #sangersequencing
Welcome to *ClevaLab* - if you like the video, please give it a 👍and subscribe for more videos. Also, if you have any questions, feel free to ask in the comments. 🤓
Loved it.. Perfect description of old methods to the new ones. Videos like this are much needed to understand these concepts visually.
I'm glad you liked it and found it helpful. 🤓 Thanks for taking the time to comment.
The animations used in this video are very helpful and well done. Excellent overview of Sanger sequencing.
Thanks for your comment. 🤓 I'm so glad you liked it.
Can't thank you enough!!!❤
Thank you so much, you can't imagine how much you have helped me. I have been confused for more an year and ths cleared everything
❤this is so good, composed & clear... Thanks
Thanks for taking the time to comment. 🤓 I'm glad you enjoyed the video.
I studied Microbiology at university from '93-'98 and this was all a black box. I appreciate the depth of this exploitation. Thank you!
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Very deatiled and informative video with excellent visuals, thank you!!!
Thanks so much! I'm glad you liked it. 🤓
Your Videos are so helpful 😊.Thanks a lot ❤
Thanks for taking the time to comment. 🤓 I'm glad you're enjoying the videos.
Best and detailed video
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Compared to so many materials, this one really made me understand it . Thanks a lot.
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This is brilliant. Thank you so much
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thabk you so much! This is so clear!
Thank you for watching. 🤓 I'm glad you liked it. 👍
wow, just a fantastic video thank you.🥰
Thanks for your comment. 🤓 I'm so glad it helped you.
Amazing video :D thank you very much 💛
I'm glad you liked it. Thanks for taking the time to comment. 🤓
Wow! You explained so well. It's easy to understand. You should be a good teacher. Thank you
I'm glad you found it useful. 🤓 Thanks for taking the time to comment.
Hey.. your videos are great. would love to see more of your videos. I am not finding many here. Please upload more videos. May be on fourth-generation technologies ;)
Thanks for your comment. 🤓 Ah yes, I'm working on more! Yes, I've got PacBio and Nanopore on my list. 👍
you make great videos, you should do one on western immunoblotting
Thanks for your comment. 🤓 I'm glad you're enjoying the videos. Good idea, I'll put that one on my list. 👍
Great video!
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This is one of the best videos I have seen in biotech so far.
Thanks so much! That's great to hear. I'm glad you enjoyed the video. 🤓
Excelent video. I just had a doubt about why exactly the radiolabel was added to the dATP. In other materials it says that the radioactive or fluorescent label is added to the ddNTPs, which makes more sense to me. But, since the dATP could be added at a random position in the chain, why did Dr. Sanger do it?
Hello, thanks for the explanation it was very very clear. Thank you also for a previous response explaining why we can be certain to get every nucleotide sequenced during the first iteration. But i just had one question about the comparison between Sanger sequencing and NGS, when you talked about sensitivity to detect a base within a background of other DNA, i didn't understand if it was a bad thing or not. Only* 15-20% implied that greater should be better or am i incorrect ?
thank you for this amazing video and for providing a pdf file with it. just I have a question, as the incorporation of ddNTPs occur RANDOMLY, how can we make sure , that each position of Adenine nucleotide for example along the template will be attached to a ddTTP once??? I mean, what if this nucleotide always with each cycle attached to a normal dTTP , this means that this nucleotide won't be read , right?
Thanks for your comment. 🤓 In the video, I did oversimplify things. For simplicity, only one strand of DNA was illustrated. However, the usual input amount for Sanger sequencing for human genomic DNA is around 200 ng. In 200 ng of human genomic DNA, there are almost 60,000 copies of the genome. This means that multiple labelled fragments are generated for each base in the sequence. The higher the input of DNA (and hence copies), the higher the final signal will be for that base in the sequencing result. There will be differences in signal intensity on the sequencing trace (chromatogram) due to the slightly different amounts of fragments at each base position. That’s why the heights of the peaks are somewhat different.
I hope this makes sense. Please let me know if you have any further questions.
@@ClevaLab yes I get it . thank you so much for this amazing explanation.😊
Very impressive animation.
@3:16 dNTPs are not radiolabeled instead ddNTPs are radiolabeled.
Thanks for your comment. 🤓 I'm glad you liked the animation.
In the original article by Sanger _et al._ the *dATPs* were radiolabeled, see the article here: www.pnas.org/doi/abs/10.1073/pnas.74.12.5463
plz upload more molecular biology videos.thanks
Thanks for your comment. 🤓 I hope to do so soon!
i don't understand why there is only one primer in Sanger sequencing TT
Thank you so much, you can't imagine how much you have helped me. I have been confused for more an year and ths cleared everything
Thanks for your comment. 🤓 I'm so glad it was helpful.