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Single-molecule super-resolution imaging of chromosomes and in situ haplotype visualization using Oligopaint FISH probes

MPG-Autoren
http://pubman.mpdl.mpg.de/cone/persons/resource/persons172959

Jungmann,  Ralf
Jungmann, Ralf / Molecular Imaging and Bionanotechnology, Max Planck Institute of Biochemistry, Max Planck Society;

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ncomms8147.pdf
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Zitation

Beliveau, B. J., Boettiger, A. N., Avendano, M. S., Jungmann, R., McCole, R. B., Joyce, E. F., et al. (2015). Single-molecule super-resolution imaging of chromosomes and in situ haplotype visualization using Oligopaint FISH probes. NATURE COMMUNICATIONS, 6: 7147. doi:10.1038/ncomms8147.


Zitierlink: http://hdl.handle.net/11858/00-001M-0000-0029-A6C5-B
Zusammenfassung
Fluorescence in situ hybridization (FISH) is a powerful single-cell technique for studying nuclear structure and organization. Here we report two advances in FISH-based imaging. We first describe the in situ visualization of single-copy regions of the genome using two single-molecule super-resolution methodologies. We then introduce a robust and reliable system that harnesses single-nucleotide polymorphisms (SNPs) to visually distinguish the maternal and paternal homologous chromosomes in mammalian and insect systems. Both of these new technologies are enabled by renewable, bioinformatically designed, oligonucleotide-based Oligopaint probes, which we augment with a strategy that uses secondary oligonucleotides (oligos) to produce and enhance fluorescent signals. These advances should substantially expand the capability to query parent-of-origin-specific chromosome positioning and gene expression on a cell-by-cell basis.