In Situ Hybridization Workflow
Step 1
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Labeling
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LabelingLabel your DNA or RNA probes and oligonucleotides using one-step direct methods, thiol-based indirect labeling, or specific 5' and 3' end tagging techniques. Carbohydrate LabelingDNA / RNA Labeling |
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Step 2
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Characterrization
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CharacterizationConfirm probe customization and label incorporation. For example, use the QuantTag Biotin Quantitation kit to determine the number of biotin molecules incorporated in each strand of nucleic acid. Affinity Binding MatricesBiotin Quantitation |
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Step 3
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Hybridization
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HybridizationOptimize the probe concentration and buffer conditions to ensure effective annealing to complementary sequences. Slide Preparation |
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Step 4
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Blocking
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BlockingReduce or eliminate unwanted background (non-specific) staining on tissue sections and cell preparations using a blocking solution. Non-specific staining can result from endogenous enzyme or tissue elements, including endogenous enzyme activity, presence of Fc receptors, or interactions of detection reagents with tissue or cell proteins and other macromolecules. Choose a blocking solution based on the results of negative control sections. Blocking Reagents |
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Step 5
| Detection
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Step 6 | Counterstain & Mount
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