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Sanger and Maxam-Gilbert Sequencing

Sanger and Maxam-Gilbert Sequencing. Cindy Wang IGSP Center for Genome Ethics, Law & Policy July 28, 2005. Sanger Method. Chain termination method Single-chain Instruments: Primer (denatured, radioactively labeled restriction enzyme) DNA Dideoxyribonucleotides (ddNTP)

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Sanger and Maxam-Gilbert Sequencing

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  1. Sanger and Maxam-Gilbert Sequencing Cindy Wang IGSP Center for Genome Ethics, Law & Policy July 28, 2005

  2. Sanger Method • Chain termination method • Single-chain • Instruments: • Primer (denatured, radioactively labeled restriction enzyme) • DNA • Dideoxyribonucleotides (ddNTP) • Dioxyribonucleotides (dNTP)

  3. Relevant History:1950’s and 1960’s • Chris Anfinsen (1954) • Radioactive labeling • Frances Crick (1962) • Conversion to sequencing • Brownlee and Barrell (1965) • Radioactive labeling of small oligonucleotides • C. Weissmann (1969) • RNA copying

  4. Relevant History: 1970’s • Nathans, Arber and Smith (1970): Restriction enzymes • Hans Kosel (1970): Synthesize DNA primers • Wu and Kaiser (1971): DNA copying • Herb Boyer (1972): EcoR1 • Sanger and Coulson (1975): Plus and minus method

  5. Citation Trends Sanger, F., Nicklen, S. & Coulson, A.R. DNA Sequencing with Chain-terminating Inhibitors. Proceedings of the National Academy of Sciences USA, 74, 5463-7.

  6. Citation Trends Sanger F & Coulson A.R. A Rapid Method for Determining Sequences in DNA by Primed Synthesis with DNA polymerase. J. Mol. Biol. 94:441-8, 1975.

  7. Patent Citation Trends Sanger, F., Nicklen, S. & Coulson, A.R. DNA Sequencing with Chain-terminating Inhibitors. Proceedings of the National Academy of Sciences USA, 74, 5463-7.

  8. Maxam-Gilbert Sequencing • Chemical cleavage • Double-stranded • Instruments: • Radioactive probe • Restriction enzymes • Nucleotide-specific enzymes

  9. Relevant History • Watson (1960) • Interest in genetics • Brookes and Lawley (1963) • Alkylation of nucleic acids • Mirzabekov (1975) • Methylation

  10. Citation Trends Maxam, A. & Gilbert, W. A New Method of Sequencing DNA. Proceedings of the National Academy of Sciences, USA, 74, 560-4 (1977).

  11. Patent Citation Trends

  12. M-G Patents • Maxam: • 1979 patent on gel electrophoresis apparatus • Gilbert: • 1982 patent on mature protein synthesis • 1983 patent on protein synthesis • 1986 patent on recombinant DNA molecule • 1990 patent on peptides in HIV pathogenesis • 1991 patent on peptides in HIV pathogenesis

  13. Analysis • Why patent/not patent? Does it matter? • Policies: Compare with PCR • Institutional environment • Political environment • Social factors • Gordon conferences • Colleagues

  14. Questions and Answers Thank you for listening!

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