YAC Protocols

1. Genomic Reconstruction by Mitotic Recombination of YACs

   • David Markie, Jiannis Ragoussis

   Pages 217-230

2. Amplification of the Copy Number of YACs

   • Lucy L. Ling, Douglas R. Smith, Donald T. Moir

   Pages 231-237

3. Transfer of YAC Clones to New Yeast Hosts

   • Forrest Spencer, Giora Simchen

   Pages 239-252

4. Use of ACEDB as a Database for YAC Library Data Management

   • Ian Dunham, Gareth Ll. Maslen

   Pages 253-280

5. YAC Transfer into Mammalian Cells by Cell Fusion

   • Nicholas P. Davies, Clare Huxley

   Pages 281-292

6. YAC Transfer by Microinjection

   • Andreas Schedl, Brenda Grimes, Lluís Montoliu

   Pages 293-306

7. Transfection of Mammalian Cells via Lipofection

   • William M. Strauss

   Pages 307-327

8. The Isolation of cDNAs by Hybridization of YACs to cDNA Libraries

   • Russell G. Snell

   Pages 329-336

9. cDNA Selection with YACs

   • Satish Parimoo

   Pages 337-358

10. Markers, Selection, and Media in YAC Cloning

    • David Markie

    Pages 359-371

11. Back Matter

    Pages 373-378

    PDF

Yeast artificial chromosomes (YACs) have their origins in the molecular genetic analysis of the yeast Saccharomyces cerevisiae. The construction of self-maintaining genetic elements from isolated frag­ ments of the yeast genome defined DNA sequences necessary for chro­ mosome function has provided telomeres, centromeres, and autonomous replicating sequences. In 1987 a reversal of the strategy put these short functional DNA sequences to work in cloning vectors, producing "yeast" chromosomes largely composed of foreign DNA. Initially the insert size of clones averaged several hundred kilobasepairs, a remarkable achieve­ ment. Rapid progress with cloning technology has since enabled the construction of YAC libraries with average insert sizes of around 1 Mb, with many clones exceeding that size, and YACs remain the largest capacity microbiological cloning system available. They effectively bridge the size gap between bacterial cloning (plasmids, cosmids, PI, and bacterial artificial chromosomes) and what could be considered mammalian cloning systems (somatic cell hybrids and irradiati- fusion gene transfer hybrids). YACs also brought with them a conceptual revolution in the man­ agement of clone libraries. The large carrying capacity of YACs, with subsequent reduction in the total number required, meant that it was conceivable to store clones individually rather than as pools that require constant re-plating. Each clone in the library has a unique address and, with successive screenings, information accumulates about individual clones.

...timely...loaded with useful tips and information.-FEBS Letters

• Paediatric Research Unit, United Medical and Dental Schools of Guy’s and St. Thomas’ Hospitals, London

  David Markie

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