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  • What are the formats of hybridization?

    Hybridization solution: The probe and target nucleic acid are in a liquid. The hybridization conditions must be in order. The rate of formation of the duplex under these conditions is high. The problem with this type of hybridization is the elimination of unreacted probe, using among other methods nuclease S1-precipitation with trichloroacetic or hybridization protection assay.

    Solid-phase Hybridization: The target nucleic acid or the probe are immobilized on filters. These can be of nitrocellulose (heat setting) or nylon (fixing by UV light). Once the attachment is added to the labeled probe will seek its complementary sequence in the target nucleic acid that we identify. The probe does not react, unstable hybrids or non-specific binding are removed by washing. Its sensitivity is lower than the hybridization solution.

    Now also has managed to fix the nucleic acids plastic microtiter plates.

    One type of solid-phase hybridizations are widely used in molecular biology: the Southern blot (DNA cut with restriction enzymes and hybridized with a probe of DNA) and Northern Blot (RNA and hybridized separately with probes of DNA or RNA). These techniques consist of an initial separation of molecules based on their molecular weight by electrophoresis, capillary transfer these molecules into a filter paper or nylon, fixing, hybridization with the probe of interest and detection.

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