Jonathan Weissman (UCSF/HHMI): DNA Sequencing
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- Опубликовано: 10 июн 2024
- www.ibiology.org/techniques/s...
In this lecture, Weissman gives an overview of the methodology that allows the sequence of DNA to be determined. He begins by explaining the classic Sanger sequencing technique using radioactively labeled nucleotides and gel electrophoresis. Next, advances such as fluorescently labeled nucleotides and capillary electrophoresis are introduced. Weissman then explains how automation and improved computing power allowed whole genomes to be sequenced, albeit slowly and at significant expense. Finally he introduces one of the "next-gen" sequencing technologies in which DNA is sequenced directly on a slide allowing millions of pieces of DNA to be sequenced in parallel. Weissman predicts that using this vastly improved technology will soon put the cost of determining an individual's genome at as little as $1000.
A fantastic review of a vast and complex subject, thank you very much indeed.
Love the progression of hand movement...
that was fantastic thank you!
very nice video, great explanations
Very nice vedeo and presentation ... thanks
Very nice video!
Thanks.. u make things clear for me.
This video was made in 2009, while we are in year 2020 now. So, which combination of nowadays technologies may eventually enable personal human genome sequencing down to clinical level (at a scale level of 1000 persons per day) and shorten the sequencing time to within 12 hours? (If cost is not a problem)
Good lecture...very interesting.. :)
very helpful thank you so much
Shouldnt the sequence be CGAT at 3: 58.. The gel would have to be read bottom to top right?
excellent, thanks! :)
How do they design the primer without knowing the fragment's seq?
in this example, they are using 5' primers, so it's back to front. but you would be correct if they were sequencing from the front end.
I am a little confused. if we use radioactive labeled primers, then the biggest band in gel will be present in all lanes. this is because the primer is labeled and consequently each band will be labeled whether is ended by a certain ddNTP or not. in other words, if in the lane of ddA, the strand has 2 T and the end of strand isnt T, then we will end up with 3 different strands. one ended at the first T, one end at the second T and one end at the end of the strand which is not T. however these 3 strands will appear in the lane of ddA. the last strand will appear in all other lanes. is this correct or not? help?
you won't have a terminating T since the reaction in the A lane has no ddT, it can never terminate at a T.
i dont understand if there is for example two Cytosine's in row if ddCTP incorporate every time in first cytosine how can we determine there is second after that?
I don't think it incorporates every time, you add enough ddCTP such that there is a possibility for incorporation
+Christopher Brown Yes! That is exactly correct.
he didn't go into details but only one nucleotide is allowed to incorporate using illumina technology due to there being a blocker added (probably part of the fluorophore). the blocker is washed away (with the previous dntps) before the next dntp is added. the number of cycles the sequencing instrument goes through is based on the read length you specify.
Please apply and build and costruct and distribute this technology to all village health clinics, public health center, public hospital clinic to make sure all civilian back to as young as 20's in good perfect health to spread more good deed among people and all earthling?
so what about dino DNA? is it possible to 73e4
no.
oy! no mention of PCR (and its semi-crazy inventor)
DNA (-) "runs to red" (+)