Chromosomal detection of simple sequence repeats (SSRs) using nondenaturing FISH (ND-FISH) |
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Authors: | Ángeles Cuadrado Nicolás Jouve |
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Institution: | 1. Department of Cell Biology and Genetics, University of Alcalá de Henares, 28871, Alcalá de Henares, Madrid, Spain
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Abstract: | Simple Sequence Repeats (SSRs) are known to be scattered and present in high number in eukaryotic genomes. We demonstrate
that dye-labeled oligodeoxyribonucleotides with repeated mono-, di-, tri, or tetranucleotide motifs (15-20 nucleotides in
length) have an unexpected ability to recognize SSR target sequences in non-denatured chromosomes. The results show that all
these probes are able to invade chromosomes, independent of the size of the repeat motif, their nucleotide sequence, or their
ability to form alternative B-DNA structures such as triplex DNA. This novel and remarkable property of binding SSR oligonucleotides
to duplex DNA targets permitted the development of a non-denaturing fluorescence in situ hybridization method that quickly
and efficiently detects SSR-enriched chromosome regions in mitotic, meiotic, and polytene chromosome spreads of different
model organisms. These results have implications for genome analysis and for investigating the roles of SSRs in chromosome
structure and function. |
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Keywords: | |
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