A versatile genome-scale PCR-based pipeline for high-definition DNA FISH

ArticleinNature Methods 10(2) · December 2012with77 Reads
Impact Factor: 32.07 · DOI: 10.1038/nmeth.2306 · Source: PubMed
Abstract

We developed a cost-effective genome-scale PCR-based method for high-definition DNA FISH (HD-FISH). We visualized gene loci with diffraction-limited resolution, chromosomes as spot clusters and single genes together with transcripts by combining HD-FISH with single-molecule RNA FISH. We provide a database of over 4.3 million primer pairs targeting the human and mouse genomes that is readily usable for rapid and flexible generation of probes.

    • "After deposition of a CTC containing sample on a suitable surface, upcoming multiplex fluorescent staining technologies in combination with fast fluorescent digital scanners are expected to facilitate CTC recognition among many white blood cells [30]. Upon marking of the location of CTCs on the scanned surface by appropriate scanner software, CTCs may in principle be selected by the software, to subsequently perform sophisticated in situ molecular assays to measure the number of specific mRNA molecules and specific genomic mutations on a single cell basis [19,20,313233. In case of a large cell-covered surface area, microfluidics approaches can be developed to perform for example a PCR assay in situ within single drops generated locally on the surface e.g., by electro-wetting [34]. "
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