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BOC Sciences offers custom FISH probe design and manufacturing services, from simple modifications to comprehensive novel designs. Our expert team is capable of designing probes that fulfill the specific needs of our customers.
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FISH (Fluorescence In Situ Hybridization) is an oligonucleotide probe using direct fluorescent group labeling or indirect labeling, such as biotin. FISH provides a method for visualizing and mapping genetic material in individual cells to validate chromosomal abnormalities and other genetic mutations.
FISH is a macromolecular recognition technique based on the complementary nature of DNA or DNA/RNA double strands. Its basic principles are shown below. (a) DNA probe and target sequence are the basic elements of FISH. (b) Prior to hybridization, DNA probe is labeled using PCR, cut-and-pan, and labeled with random primer. Two labeling strategies, indirect labeling (left panel) and direct labeling (right panel) are commonly used. Modified nucleotide-labeled probes containing semi-antigens are used for indirect labeling, while direct labeling adapts directly modified nucleotides containing fluorophores. (c-d) Both labeled probes and target DNA apply the denatured-annealed-denatured process to hybridize target DNA and nucleic acid probe. (e) If probe is indirectly labeled, an additional step is required to visualize the non-fluorescent semi-antigen with an enzymatic or immunological detection system. The hybrid between target gene and probe is formed according to the principle of base complementarity, and target signal is visualized by fluorescence microscopy. This allows for the determination of relative qualitative, quantitative and localization analysis of the nucleic acid sequence.
Fig.1 The principle of fluorescence in situ hybridization (FISH). (Michael R., 2005)
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