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This protocol describes the synthesis of an RNA probe labeled with digoxigenin (DIG) for use in whole mount in situ hybridization (WISH). The procedure involves digesting plasmid DNA with the appropriate restriction enzyme, extracting the DNA with phenol–chloroform, and precipitating it with ethanol (EtOH) or purifying it using a column. The purified DNA is then used as a template for RNA synthesis. The final RNA product is labeled with DIG for subsequent detection in WISH assays.

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Synthesis of RNA–Digoxigenin Probe

  • 实验方案

  • 发表日期：

    2025年

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  • Karuppusamy Arunachalam,
  • Puthanpura Sasidharan Sreeja

摘要

This protocol describes the synthesis of an RNA probe labeled with digoxigenin (DIG) for use in whole mount in situ hybridization (WISH). The procedure involves digesting plasmid DNA with the appropriate restriction enzyme, extracting the DNA with phenol–chloroform, and precipitating it with ethanol (EtOH) or purifying it using a column. The purified DNA is then used as a template for RNA synthesis. The final RNA product is labeled with DIG for subsequent detection in WISH assays.