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New polymerases for old DNA: molecular breeding of polymerases for damage bypass and ancient DNA amplification

New polymerases for old DNA: molecular breeding of polymerases for damage bypass and ancient DNA amplification. Philipp Holliger MRC Laboratory of Molecular Biology. The “unreadable” scrolls of Herculaneum. Herculaneum. Mt. Vesuvius (Naples, Italy). Villa dei Papyrii. ancient text.

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New polymerases for old DNA: molecular breeding of polymerases for damage bypass and ancient DNA amplification

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  1. New polymerases for old DNA:molecular breeding of polymerases for damage bypass and ancient DNA amplification Philipp Holliger MRC Laboratory of Molecular Biology

  2. The “unreadable” scrolls of Herculaneum Herculaneum Mt. Vesuvius (Naples, Italy) Villa dei Papyrii ancient text Charcoal? ancient scroll

  3. Ancient DNA Neanderthal man (Homo neanderthaliensis) our closest extinct relative PCR (1993 Nobel prize in Chemistry)

  4. Damage found in ancient DNA From Hofreiter et al Nat Rev Genet. 2001 2:353

  5. The directed evolution cycle

  6. Oil phase Water droplet Ghadessy et al (2001) PNAS, 98, 4552

  7. Molecular breeding Crameri et al (1998) Nature, 391, 288 Zhao et al (1998) Nature Biotechnology, 16,258

  8. Selection strategy d’Abbadie et al (2007) Nature Biotechnology, 25, 939

  9. Non-cognate primer-template structures d’Abbadie et al (2007) Nature Biotechnology, 25, 939

  10. Damage bypass d’Abbadie et al (2007) Nature Biotechnology, 25, 939

  11. PCR amplification of DNA containing two abasic sites Taq Tth Tfl 3A10 d’Abbadie et al (2007) Nature Biotechnology, 25, 939

  12. PCR amplification of ancient cave bear DNA d’Abbadie et al (2007) Nature Biotechnology, 25, 939

  13. PCR amplification of ancient cave bear DNA d’Abbadie et al (2007) Nature Biotechnology, 25, 939

  14. Ice age genomics Cave bear (Ursus spelaeus) Neanderthal man (Homo neanderthaliensis) 106 bp sequenced: Green et al Nature (2006) 444, 330; Noonan et al Science (2006) 314, 1113 Noonan et al, Science (2005), 309, 597 Wolly mammoth (Mammuthus primigenius) Mt genome (Krause et al Nature (2006),439, 724; Poinar et al Science (2006) 311, 392)

  15. Acknowledgements Max Planck Institute for Molecular Anthropology Michael Hofreiter Svante Pääbo NIH Section on DNA Replication, Repair and Mutagenesis Alexandra Vaisman Roger Woodgate MRC Laboratory of Molecular Biology Marc d’Abbadie Farid J. Ghadessy David Loakes

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