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ISH Probes Derived from BACs, Including Microwave Treatment for Better FISH Results

  • Anja WeiseEmail author
  • Kristin Mrasek
  • Nadezda Kosyakova
  • Hasmik Mkrtchyan
  • Madeleine Gross
  • Vivien Klaschka
  • Thomas Liehr
Protocol
Part of the Springer Protocols Handbooks book series (SPH)

Besides the well-known applications of bacterial artificial chromosomes (BACs) in classical molecular genetics, BACs are also used for molecular cytogenetic studies. BACs, as well as other locus-specific probes like cDNA, plasmids, cosmids, fosmids, P1-clones or yeast artificial chromosomes (YACs), can be labeled with fluorochromes and applied in FISH experiments. Various applications are possible, like gene mapping, FISH banding, determination of chromosomal breakpoints, characterization of derivative chromosomes, studies of interphase architecture, and karyotypic evolution studies. Here the basic principle of hybridizing BACs in situ on chromosome preparations is outlined. Moreover, an overview of possible issues that can be studied using BACs as FISH probes is provided. Finally, a shortened and more efficient FISH protocol using microwave treatment which yields results that can be evaluated within a few hours is presented.

Keywords

Bacterial Artificial Chromosome Microwave Treatment Yeast Artificial Chromosome Chromosome Painting Probe Partial Chromosome Painting 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

Acknowledgments Supported in part by the Evangelische Studienwerk e.V. Villigst, IZKF Jena (Start-up S16, TP 3.7), Friedrich Schiller University Jena, TMWFK (B307–04004), Stiftung Leukämie and Stefan-Morsch-Stiftung.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2009

Authors and Affiliations

  • Anja Weise
    • 1
    Email author
  • Kristin Mrasek
    • 1
  • Nadezda Kosyakova
    • 1
  • Hasmik Mkrtchyan
    • 2
  • Madeleine Gross
    • 1
  • Vivien Klaschka
    • 1
  • Thomas Liehr
    • 1
  1. 1.Institut für Humangenetik und AnthropologieJenaGermany
  2. 2.Department of Genetic and Laboratory of CytogeneticsState UniversityJerewanArmenia

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