What Is Sanger Sequencing?
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- เผยแพร่เมื่อ 24 ส.ค. 2020
- Learn about a workhorse technique of lab research since the late 1970's.
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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
Wow. Amazing explanation, it really helped me understand how genius this method is.
Very concise yet gets the concept across in an excellent manner which is very easy to understand! Thank you!
Thank you! You're a lifesaver!
Great explanation
Great thanx
Finally a good video
It's between anger and sadness actually, lol.
wow
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
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
@@user-nf2in4fs5u 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.
Huh ?
It still doesn't explain exactly how it's done.