Detection of short repeated genomic sequences on metaphase chromosomes using padlock probes and target primed rolling circle DNA synthesis |
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Authors: | Jakob S Lohmann Magnus Stougaard Jørn Koch |
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Institution: | (1) Institute of Pathology, Aarhus University, 8000 Aarhus, C, Denmark |
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Abstract: | Background In situ detection of short sequence elements in genomic DNA requires short probes with high molecular resolution and powerful
specific signal amplification. Padlock probes can differentiate single base variations. Ligated padlock probes can be amplified
in situ by rolling circle DNA synthesis and detected by fluorescence microscopy, thus enhancing PRINS type reactions, where
localized DNA synthesis reports on the position of hybridization targets, to potentially reveal the binding of single oligonucleotide-size
probe molecules. Such a system has been presented for the detection of mitochondrial DNA in fixed cells, whereas attempts
to apply rolling circle detection to metaphase chromosomes have previously failed, according to the literature. |
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