sanger sequencing. 1) get some dna to sequence *le pick… *le smash… *le precipitate… *le dna...
TRANSCRIPT
![Page 1: Sanger Sequencing. 1) Get Some DNA To Sequence *le pick… *le smash… *le precipitate… *le DNA in le solution](https://reader035.vdocuments.us/reader035/viewer/2022062314/56649d9d5503460f94a86836/html5/thumbnails/1.jpg)
Sanger Sequencing
![Page 2: Sanger Sequencing. 1) Get Some DNA To Sequence *le pick… *le smash… *le precipitate… *le DNA in le solution](https://reader035.vdocuments.us/reader035/viewer/2022062314/56649d9d5503460f94a86836/html5/thumbnails/2.jpg)
1) Get Some DNA To Sequence
*le pick…
*le smash…
*le precipitate…*le DNA in le solution
![Page 3: Sanger Sequencing. 1) Get Some DNA To Sequence *le pick… *le smash… *le precipitate… *le DNA in le solution](https://reader035.vdocuments.us/reader035/viewer/2022062314/56649d9d5503460f94a86836/html5/thumbnails/3.jpg)
2) Add a Primer That Binds To Area You Want to Sequence
*le DNA in le solution * le primer
Base pair with gene to sequence
3’ end for adding nucleotides
![Page 4: Sanger Sequencing. 1) Get Some DNA To Sequence *le pick… *le smash… *le precipitate… *le DNA in le solution](https://reader035.vdocuments.us/reader035/viewer/2022062314/56649d9d5503460f94a86836/html5/thumbnails/4.jpg)
3) Make Many Copies of the Gene
*le DNA + le Primers
*le DNA Polymerase
*le free nucleotides(dNTPS)
Heating Cycle:1) 94oC to break H-bonds2) 65oC Primers anneal to ssDNA3) 72oC DNA Polymerase copies genes
using dNTPS4) Repeat Many Times
![Page 5: Sanger Sequencing. 1) Get Some DNA To Sequence *le pick… *le smash… *le precipitate… *le DNA in le solution](https://reader035.vdocuments.us/reader035/viewer/2022062314/56649d9d5503460f94a86836/html5/thumbnails/5.jpg)
How Heating Cycle WorksHeating Cycle:
1) 94oC to break H-bonds2) 65oC Primers anneal to ssDNA3) 72oC DNA Polymerase copies
genes using dNTPS4) Repeat Many Times
*le DNA*le H-bonds
![Page 6: Sanger Sequencing. 1) Get Some DNA To Sequence *le pick… *le smash… *le precipitate… *le DNA in le solution](https://reader035.vdocuments.us/reader035/viewer/2022062314/56649d9d5503460f94a86836/html5/thumbnails/6.jpg)
How Heating Cycle WorksHeating Cycle:
1) 94oC to break H-bonds2) 65oC Primers anneal to ssDNA3) 72oC DNA Polymerase copies genes
using dNTPS4) Repeat Many Times
![Page 7: Sanger Sequencing. 1) Get Some DNA To Sequence *le pick… *le smash… *le precipitate… *le DNA in le solution](https://reader035.vdocuments.us/reader035/viewer/2022062314/56649d9d5503460f94a86836/html5/thumbnails/7.jpg)
4) Copy Gene Using Doubly Deoxidised, Florescent NTPs
*le many copies of gene + primers
*le DNA Polymerase
*le free nucleotides(dNTPS)
Fluorescent dye (A=Red, C= Blue…)
Doubly deoxidised (no 3’ oxygen to form
next bond)
![Page 8: Sanger Sequencing. 1) Get Some DNA To Sequence *le pick… *le smash… *le precipitate… *le DNA in le solution](https://reader035.vdocuments.us/reader035/viewer/2022062314/56649d9d5503460f94a86836/html5/thumbnails/8.jpg)
How ddNTPs WorkHeating Cycle:
1) 94oC to break H-bonds2) 65oC Primers anneal to ssDNA3) 72oC DNA Polymerase copies
genes using dNTPS4) Repeat Many Times
*le DNA*le H-bonds
![Page 9: Sanger Sequencing. 1) Get Some DNA To Sequence *le pick… *le smash… *le precipitate… *le DNA in le solution](https://reader035.vdocuments.us/reader035/viewer/2022062314/56649d9d5503460f94a86836/html5/thumbnails/9.jpg)
How Heating Cycle WorksHeating Cycle:
1) 94oC to break H-bonds2) 65oC Primers anneal to ssDNA3) 72oC DNA Polymerase copies genes
using dNTPS4) Repeat Many Times
![Page 10: Sanger Sequencing. 1) Get Some DNA To Sequence *le pick… *le smash… *le precipitate… *le DNA in le solution](https://reader035.vdocuments.us/reader035/viewer/2022062314/56649d9d5503460f94a86836/html5/thumbnails/10.jpg)
5) Repeat Many Times To Get Copies of Different Lengths all Ending in a ddNTP
*le messy mess
![Page 11: Sanger Sequencing. 1) Get Some DNA To Sequence *le pick… *le smash… *le precipitate… *le DNA in le solution](https://reader035.vdocuments.us/reader035/viewer/2022062314/56649d9d5503460f94a86836/html5/thumbnails/11.jpg)
6) Sort DNA Segments By Size With Gel Electrophoresis*le not messy mess
*le agarose gel
*le shockie shockie
![Page 12: Sanger Sequencing. 1) Get Some DNA To Sequence *le pick… *le smash… *le precipitate… *le DNA in le solution](https://reader035.vdocuments.us/reader035/viewer/2022062314/56649d9d5503460f94a86836/html5/thumbnails/12.jpg)
7) Use A Laser To “Read” Sections and Sequence DNA
*le cool laser
*le sequence