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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. 🤓 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
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.
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?
Amazing video, I got to know about many things yhrough this. I am currently trying to get to know about how to write research paper and for that I chose a topic of next generation sequencing can you provide me a pdf for that purpose?
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 ?
Thanks for your comment. 🤓 There's only one primer used in sequencing because we only want to know the sequence of one of the strands. Two primers are used in PCR because we want to create twice as much DNA each cycle. Watching the ClevaLab PCR video may make it clearer to you. Watch it here: th-cam.com/video/rpLSvEbOmqc/w-d-xo.htmlsi=PE8FQXA38oiKHL1l
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.
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
I studied Microbiology at university from '93-'98 and this was all a black box. I appreciate the depth of this exploitation. Thank you!
Thanks for your comment. 🤓 I'm glad you liked the video.
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.
Compared to so many materials, this one really made me understand it . Thanks a lot.
I'm glad you liked it. 🤓 Thanks for your comment.
very detailed yet crisp explanation ..love your videos
Thanks for your comment. 🤓 It's great to hear you find the videos helpful.
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.
Great video!
Thanks for your comment. 🤓 I'm glad it helped.
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. 🤓
Great video. It was very easy to understand the concepts due to the animations 👍👍😄
I'm so glad you found it helpful and liked the animations. Thanks for your comment. 🤓
Excellent video! Thanks
I'm glad glad you liked it. 🤓 Thanks for your comment.
thabk you so much! This is so clear!
Thank you for watching. 🤓 I'm glad you liked it. 👍
This is brilliant. Thank you so much
Thanks for taking the time to comment. 🤓 I'm glad you're enjoying the videos.
plz upload more molecular biology videos.thanks
Thanks for your comment. 🤓 I hope to do so soon!
Best and detailed video
Thanks for your comment. 🤓 I'm glad you liked it.
❤this is so good, composed & clear... Thanks
Thanks for taking the time to comment. 🤓 I'm glad you enjoyed the video.
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. 👍
wow, just a fantastic video thank you.🥰
Thanks for your comment. 🤓 I'm so glad it helped you.
Very deatiled and informative video with excellent visuals, thank you!!!
Thanks so much! I'm glad you liked it. 🤓
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
Your Videos are so helpful 😊.Thanks a lot ❤
Thanks for taking the time to comment. 🤓 I'm glad you're enjoying the videos.
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.😊
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?
Can't thank you enough!!!❤
You're welcome. 🤓
Amazing video :D thank you very much 💛
I'm glad you liked it. Thanks for taking the time to comment. 🤓
Amazing video, I got to know about many things yhrough this. I am currently trying to get to know about how to write research paper and for that I chose a topic of next generation sequencing can you provide me a pdf for that purpose?
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 ?
i don't understand why there is only one primer in Sanger sequencing TT
Thanks for your comment. 🤓 There's only one primer used in sequencing because we only want to know the sequence of one of the strands. Two primers are used in PCR because we want to create twice as much DNA each cycle.
Watching the ClevaLab PCR video may make it clearer to you. Watch it here: th-cam.com/video/rpLSvEbOmqc/w-d-xo.htmlsi=PE8FQXA38oiKHL1l
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. 👍
might be the first time i hear it referred to as the oxygen, instead of the hydroxide
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.