An improved method for generating BAC DNA suitable for FISH

J. Roohi, M. Cammer, C. Montagna, E. Hatchwell

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

Abstract

Fluorescence in situ hybridization (FISH) is commonly used to identify chromosomal aberrations such as translocations, deletions, duplications, gene fusions, and aneuploidies. It relies on the hybridization of fluorescently labeled DNA probes onto denatured metaphase chromosomes or interphase nuclei. These probes are often generated from DNA sequences cloned within bacterial artificial chromosomes (BACs). Growing these BACs in adequate amounts for FISH can be demanding. We describe FISH performed with bacteriophage Phi29 DNA polymerase amplified BAC DNA. Generating this material required significantly smaller cultures and less time than standard methods. The FISH results obtained were comparable with those obtained from standard BAC DNA. We believe this method of BAC DNA generation is useful for the entire FISH community as it improves considerably on prior methods.

Original languageEnglish (US)
Pages (from-to)7-9
Number of pages3
JournalCytogenetic and Genome Research
Volume121
Issue number1
DOIs
StatePublished - Jun 2008

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Genetics(clinical)

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