Molecular Definition of High-resolution Multicolor Banding Probes: First Within the Human DNA Sequence Anchored FISH Banding Probe Set

Abstract

(NK) S U M M A R Y Fluorescence in situ hybridization (FISH) banding approaches are standard for the exact characterization of simple, complex, and even cryptic chromosomal aberrations within the human genome. The most frequently applied FISH banding technique is the multicolor banding approach, also abbreviated as m-band, MCB, or in its whole genomic variant multitude MCB (mMCB). MCB allows the differentiation of chromosome region-specific areas at the GTG band and sub-band level and is based on region-specific microdissection libraries, producing changing fluorescence intensity ratios along the chromosomes. The latter are used to assign different pseudocolors to specific chromosomal regions. Here we present the first bacterial artificial chromosome (BAC) array comparative genomic hybridization (aCGH) mapped, comprehensive, genome-wide human MCB probe set. All 169 region-specific microdissection libraries were characterized in detail for their size and the regions of overlap. In summary, the unique possibilities of the MCB technique to characterize chromosomal breakpoints in one FISH experiment are now complemented by the feature of being anchored within the human DNA sequence at the BAC level. (J Histochem Cytochem 56:487-493, 2008)

Case Report Identification of a de novo inv dup(X)(pter→ q22) by multicolor banding in a girl with Turner syndrome

“…The color bands have a great value in identifying chromosomal abnormalities, particularly complex chromosome rearrangements, and intrachromosome exchanges (i.e., inversions, deletions, duplications, and insertions). These abnormalities cannot be easily defined by conventional cytogenetic analysis or chromosome paint (Hu et al, 2006;Weise et al, 2008). In our case the use of MCB enables an appreciation, at first glance, that X(pter→q22) regions are duplicated and inverted, and that another part of Xq is deleted.…”

High-throughput sequencing of microdissected chromosomal regions

“…Smaller parts can also be microdissected, as was previously shown for microdissection libraries with band-resolutions that were created for chromosome painting. 24 Such adaptation might be useful for specific questions. Here, we wanted to cover complete regions, including centromeric regions and repeats.…”

“…Microdissection was performed as described. 24 For amplification, we used an adapted degenerate oligonucleotide-primed PCR (DOP-PCR). 24,25 A detailed protocol for microdissection and DOP-PCR is provided in the supplementary material.…”

“…24 For amplification, we used an adapted degenerate oligonucleotide-primed PCR (DOP-PCR). 24,25 A detailed protocol for microdissection and DOP-PCR is provided in the supplementary material. Before preparing the 454 library, we verified the specificity of the microdissected material by dye-labeling an aliquot of DOP-PCR product and subsequent hybridization on metaphase chromosomes (reverse painting, reverse fluorescence in situ hybridization, FISH).…”

Preferred co-localization of chromosome 8 and 21 in myeloid bone marrow cells detected by three dimensional molecular cytogenetics

“…Multicolor banding (MCB) probe sets for chromosomes 8 and 21 (35,36) were applied in suspension-FISH (S-FISH) as previously reported (17,20,21,37). Images of 3D-preserved interphase nuclei were captured on a Zeiss Axioplan microscope and analyzed by Cell-P (Olympus) software.…”