See how the ProDye™ Sanger Sequencing System is used on the Spectrum Compact CE System in a proof-of-concept workflow for environmental microbial monitoring programs that require species-level identification: bit.ly/3Pjvhwm 👉Helpful Links Subscribe to Promega on RUclips: bit.ly/2YtQhtS More resources on our website: bit.ly/3BFhFna Link to product page: bit.ly/3av8PMX
What is Sanger sequencing? A conventional method used since 1977, is still used today In this we denature the DNA, by increasing the temperature. Cool down to cause primers to anneal. DNA polymerase then extend the DNA fragments until a fluorescently labeled dideoxyribonucleotide is reached. The chain is terminated. The colour of the florescent indicates the base at which the chain is terminated. When the DNA fragments are separated by size, we can determine the sequence of terminal nucleotides and determine the sequence of DNA/genome
@0:48 ... is missing a hydroxyl group which allows the next nucleotide to bind - you mean it does not allow the next nucleotide to bind, thus ending in termination...
Yeah, basically we need a hydroxyl group at 3' carbon atom in a pentose, for phosphodiester bond to be formed and for DNA sequencing to happen, right.. so ddNTP lacks that 3' OH group as well. So no further bond formation, and no more sequencing.
When we terminate the chain with ddNTP’s we understand that in this spesific place there is spesific nucleotide that we added in the tube and after that you repeat the action and figure out all of different spesific places too. I hope i could explain you
Not quite, in PCR our goal is to make lots of copies of DNA fragments and then analyse them in order to find the sequence we need, but with sequencing we aim for the nucleotide sequence in general, which is a bit more detailed thing that is why we use fluorescent dideoxynucleotides, hope that helped ❤
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See how the ProDye™ Sanger Sequencing System is used on the Spectrum Compact CE System in a proof-of-concept workflow for environmental microbial monitoring programs that require species-level identification: bit.ly/3Pjvhwm
👉Helpful Links
Subscribe to Promega on RUclips: bit.ly/2YtQhtS
More resources on our website: bit.ly/3BFhFna
Link to product page: bit.ly/3av8PMX
What is Sanger sequencing?
A conventional method used since 1977, is still used today
In this we denature the DNA, by increasing the temperature. Cool down to cause primers to anneal. DNA polymerase then extend the DNA fragments until a fluorescently labeled dideoxyribonucleotide is reached. The chain is terminated.
The colour of the florescent indicates the base at which the chain is terminated.
When the DNA fragments are separated by size, we can determine the sequence of terminal nucleotides and determine the sequence of DNA/genome
Spectacular explanation! It’s amazing how you simplified it thank you
Very concise yet gets the concept across in an excellent manner which is very easy to understand! Thank you!
Wow. Amazing explanation, it really helped me understand how genius this method is.
Great video! One thing--I think it's misleading to say the ddNTP is "randomly" inserted. It's inserted according to Watson-Crick rules.
He means that, by chance, a ddNTP is inserted instead of a normal dNTP, which would not stop elongation.
Yep, I get that! Just pointing out that his statement could be misinterpreted. @@poldi7
@@EllieBrowne-z1i Yeah, that could be true. You‘re right
@0:48 ... is missing a hydroxyl group which allows the next nucleotide to bind - you mean it does not allow the next nucleotide to bind, thus ending in termination...
Hi there, thanks for your comment. The dideoxynucleotide is what terminates the chain.
Hydroxyl group allows the next nucleotide to bind, thus ddNTP (which is missing one) terminates the sequence, he explained it fine.
Yeah, basically we need a hydroxyl group at 3' carbon atom in a pentose, for phosphodiester bond to be formed and for DNA sequencing to happen, right.. so ddNTP lacks that 3' OH group as well. So no further bond formation, and no more sequencing.
Great explanation
Lovely and simple to understand!
I don't understand how we get to know about the sequence and why we are terminating it with ddNTPs??
Can someone help me with this?
When we terminate the chain with ddNTP’s we understand that in this spesific place there is spesific nucleotide that we added in the tube and after that you repeat the action and figure out all of different spesific places too. I hope i could explain you
Thanks a lot! This was easy to understand
Thank you
superb explanation
Thank you! You're a lifesaver!
Finally a good video
does anyone know the name of the background music?
Fairy Meeting by Ricardo Aguero
Wait.. isn't this the procedure for Polymerase Chain reaction?
Not quite, in PCR our goal is to make lots of copies of DNA fragments and then analyse them in order to find the sequence we need, but with sequencing we aim for the nucleotide sequence in general, which is a bit more detailed thing that is why we use fluorescent dideoxynucleotides, hope that helped ❤
Great thanx
It still doesn't explain exactly how it's done.
Omg Wow i totally get it now..
I now understand why that guy left the human genome project to do it himself
Seems similar to pcr
Smith Parkway
wow
Huh ?
📚🇧🇷🙏
hiçbir şey anlamadım şimdi bir daha izleyeceğim
Woooooooooooooooooooowwww
:)