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  • Array painting: a protocol for the rapid analysis of aberrant chromosomes using DNA microarrays.

Array painting: a protocol for the rapid analysis of aberrant chromosomes using DNA microarrays.

Nature protocols (2009-11-07)
Susan M Gribble, Bee Ling Ng, Elena Prigmore, Tomas Fitzgerald, Nigel P Carter
ABSTRACT

Array painting is a technique that uses microarray technology to rapidly map chromosome translocation breakpoints. Previous methods to map translocation breakpoints have used fluorescence in situ hybridization (FISH) and have consequently been labor-intensive, time-consuming and restricted to the low breakpoint resolution imposed by the use of metaphase chromosomes. Array painting combines the isolation of derivative chromosomes (chromosomes with translocations) and high-resolution microarray analysis to refine the genomic location of translocation breakpoints in a single experiment. In this protocol, we describe array painting by isolation of derivative chromosomes using a MoFlo flow sorter, amplification of these derivatives using whole-genome amplification and hybridization onto commercially available oligonucleotide microarrays. Although the sorting of derivative chromosomes is a specialized procedure requiring sophisticated equipment, the amplification, labeling and hybridization of DNA is straightforward, robust and can be completed within 1 week. The protocol described produces good quality data; however, array painting is equally achievable using any combination of the available alternative methodologies for chromosome isolation, amplification and hybridization.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
GenomePlex® Complete Whole Genome Amplification (WGA) Kit, Optimized kit with enzyme for amplifying a variety of DNA including FFPE tissue
Sigma-Aldrich
Spermidine trihydrochloride, ≥98% (TLC)
Sigma-Aldrich
Sodium citrate tribasic dihydrate, ACS reagent, ≥99.0%
Sigma-Aldrich
Griess reagent (modified)