1 / 10

Deciphering DNA: Pioneering Gene Sequencing Techniques

Explore the evolution of gene sequencing, from Watson & Crick's discovery of DNA double helix to Sanger's chain termination method and subsequent advancements. Learn about early sequencing efforts, nucleic acid structure, and key breakthroughs by scientists like Holley, Wu, and Fiers. Discover how techniques like Southern Blotting, restriction enzymes, and Sanger's dideoxynucleotide method revolutionized genetic research. References to influential publications and Nobel laureates in chemistry underscore the significance of these milestones.

anitahall
Download Presentation

Deciphering DNA: Pioneering Gene Sequencing Techniques

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. First generation Gene Sequencing Jessie Marlenee BIOL 546 https://en.wikipedia.org/wiki/Phi_X_174

  2. Recap • Watson & Crick discover DNA double helix 1953 • Restriction Enzymes • Southern Blotting • Cloning https://en.wikipedia.org/wiki/Nucleic_acid_double_helix https://en.wikipedia.org/wiki/Restriction_enzyme

  3. Initial sequencing efforts • Proteins • RNA • Holley and colleagues (1965) • alanine tRNA from Saccharomyces cerevisiae • Wu and Kaiser (1968-71) • Incorporation of radiolabeled nucleotides into strand to determine sequence • Walter Fiers' laboratory (1972-76) • Coat protein of bacteriophage MS2, followed by complete genome four years later (3569 nucleotides of ssRNA) • Replacement of 2D fractionation https://en.wikipedia.org/wiki/Bacteriophage_MS2 Kaiser, A. D. and Wu, R. 1968.

  4. Sanger’s “Plus and Minus” technique(1975) https://en.wikipedia.org/wiki/Phi_X_174 https://slideplayer.com/slide/5188821/

  5. Maxam and Gilbert’s Chemical cleavage technique (1977) Matthews and van Holde: Biochemistry, 2nd edition

  6. Sanger’s “chain termination” technique (1977) Also known as the dideoxynucleotide method or sanger sequencing

  7. 1980 Nobel prize in chemistry • ½ Awarded to Paul Berg • ¼ Awarded to Walter Gilbert • ¼ Awarded to Frederick Sanger

  8. References • Franca, L. T., Carrilho, E., and Kist, T. B. 2002. A review of DNA sequencing techniques. Q. Rev. Biophys. 35:169–200. • Guzvic, M. 2013. The history of DNA sequencing. J Med Biochem. 32:301–12. • Heather, J. M., and Chain, B. 2016. The sequence of sequencers: The history of sequencing DNA, Genomics. 107:1-8. • Hutchison C. A. III. 2007. DNA sequencing: bench to bedside and beyond. Nucleic Acids Res. 35:6227–6237. • Holley, R. W., Apgar, J., Everett, G. A., Madison, J. T., Marquisee, M., Merrill, S. H., Penswick, J. R. and Zamir, A. 1965. Structure of a ribonucleic acid. Science. 147:1462–1465. • Kaiser, A. D. and Wu, R. 1968. Structure and function of DNA cohesive ends. Cold Spring Harb. Symp. Quant. Biol. 33:729–734. • Mardis, E. R. 2013. Next-generation sequencing platforms. Annu. Rev. Anal. Chem. 6:287–303. • Maxam, A. M. & Gilbert, W. 1977. A new method for sequencing DNA. Proc. Natn. Acad. Sci. 74:560–564. • Sanger, F. and Coulson, A. R. 1975. A rapid method for determining sequences in DNA by primed synthesis with DNA polymerase. J. Mol. Biol., 94:441–448. • Sanger, F., Nicklen, S. & Coulson, A. R. 1977. DNA sequencing with chain-terminating inhibitors. Proc. Natn. Acad. Sci. 74:5463–5467. • The Nobel Prize in Chemistry 1980. NobelPrize.org. Nobel Media AB 2018. Thu. 13 Sep 2018. https://www.nobelprize.org/prizes/chemistry/1980/summary/ • Wu, R. and Kaiser, A. D. 1968. Structure and base sequence in the cohesive ends of bacteriophage lambda DNA. J. Mol. Biol., 35:523–537.

  9. Questions?

More Related