Rapid detection of genomic imbalances using micro-arrays consisting of pooled BACs covering all human chromosome arms

Nucleic Acids Res. 2005 Oct 12;33(18):e159. doi: 10.1093/nar/gni161.

Abstract

A strategy is presented to select, pool and spot human BAC clones on an array in such a way that each spot contains five well performing BAC clones, covering one chromosome arm. A mini-array of 240 spots was prepared representing all human chromosome arms in a 5-fold as well as some controls, and used for comparative genomic hybridization (CGH) of 10 cell lines with aneusomies frequently found in clinical cytogenetics and oncology. Spot-to-spot variation within five replicates was below 6% and all expected abnormalities were detected 100% correctly. Sensitivity was such that replacing one BAC clone in a given spot of five by a BAC clone from another chromosome, thus resulting in a change in ratio of 20%, was reproducibly detected. Incubation time of the mini-array was varied and the fluorescently labelled target DNA was diluted. Typically, aneusomies could be detected using 30 ng of non-amplified random primed labelled DNA amounts in a 4 h hybridization reaction. Potential application of these mini-arrays for genomic profiling of disseminated tumour cells or of blastomeres for preimplantation genetic diagnosis, using specially designed DNA amplification methods, are discussed.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aneuploidy*
  • Cell Line
  • Chromosomes, Artificial, Bacterial*
  • Chromosomes, Human*
  • Female
  • Genomics / methods*
  • Humans
  • Male
  • Oligonucleotide Array Sequence Analysis / methods*
  • Time Factors