LNA-modified oligonucleotides are highly efficient as FISH probes.
Fluorescence in situ hybridization (FISH) is a highly useful technique with a wide range of applications including the delineation of complex karyotypes, prenatal diagnosis of aneuploidies, screening for diagnostic or prognostic markers in cancer cells, gene mapping and gene expression studies. However, it is still a fairly time-consuming method with limitations in both sensitivity and resolution. Locked Nucleic Acids (LNAs) constitute a novel class of RNA analogs that have an exceptionally high affinity towards complementary DNA and RNA. Substitution of DNA oligonucleotide probes with LNA has shown to significantly increase their thermal duplex stability as well as to improve the discrimination between perfectly matched and mismatched target nucleic acids. To exploit the improved hybridization properties of LNA oligonucleotides in FISH, we have designed several LNA substituted oligonucleotide probes specific to different human-specific repetitive elements, such as the classical satellite-2, telomere and alpha-satellite repeats. In the present study we show that LNA modified oligonucleotides are excellent probes in FISH, combining high binding affinity with short hybridization time.
Wilhelm Johannsen Centre for Functional Genome Research, Dept. of Medical Genetics, Inst. of Medical Biochemistry and Genetics, The Panum Institute, University of Copenhagen, Denmark. asli@medgen.ku.dk
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Silahtaroglu
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Pfundheller
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Koshkin
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Tommerup
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Kauppinen
D002467:Cell Nucleus / Q000235:genetics; D002478:Cells, Cultured; D002503:Centromere / Q000235:genetics; D002877:Chromosomes, Human / Q000235:genetics; D015342:DNA Probes / Q000235:genetics*; D004276:DNA, Satellite / Q000235:genetics; D006570:Heterochromatin / Q000235:genetics; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007399:Interphase / Q000235:genetics; D008214:Lymphocytes / Q000737:chemistry / Q000378:metabolism; D008677:Metaphase / Q000235:genetics; D015394:Molecular Structure; D009841:Oligonucleotides / Q000235:genetics*; D016376:Oligonucleotides, Antisense / Q000235:genetics*; D015347:RNA Probes / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics; D016615:Telomere / Q000235:genetics
D015342:DNA Probes; D004276:DNA, Satellite; D006570:Heterochromatin; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense; D015347:RNA Probes; C477371:locked nucleic acid