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Related Experiment Videos

Loading and transfer control for Northern hybridization.

R Correa-Rotter1, C N Mariash, M E Rosenberg

  • 1Department of Medicine, University of Minnesota, Minneapolis 55455.

Biotechniques
|February 1, 1992
PubMed
Summary
This summary is machine-generated.

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This study introduces a cost-effective RNA analysis method using computer-assisted video densitometry. It accurately quantifies RNA loading, transfer, and integrity for Northern hybridization without needing housekeeping genes.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Northern hybridization is a key technique for RNA analysis.
  • Accurate quantification of RNA loading and integrity is crucial for reliable results.
  • Existing methods often rely on variable housekeeping genes for normalization.

Purpose of the Study:

  • To develop a simple, inexpensive, and reliable method for RNA quantification and integrity assessment.
  • To provide an alternative to housekeeping gene normalization in Northern hybridization.

Main Methods:

  • Utilizing computer-assisted video densitometry on negative photographs.
  • Quantifying ethidium bromide-stained 28S and 18S ribosomal RNA (rRNA) fixed to nylon membranes.
  • Assessing RNA loading, transfer efficiency, and integrity.

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Main Results:

  • The developed method provides accurate quantification of RNA loading and transfer.
  • It effectively assesses RNA integrity.
  • Eliminates the need for quantifying "housekeeping" genes, reducing variability.

Conclusions:

  • This technique offers a straightforward and economical approach to RNA analysis for Northern hybridization.
  • It enhances the reliability of RNA quantification by avoiding normalization issues.
  • The method is suitable for routine laboratory use.