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The RNase Protection Assay.

M H Finer1

  • 1Cell Genesys Inc., Foster City, CA.

Methods in Molecular Biology (Clifton, N.J.)
|March 19, 2011
PubMed
Summary
This summary is machine-generated.

Quantifying mRNA in gene transfer studies is improved with mRNA protection assays using labeled probes. RNA probes offer an efficient alternative to DNA probes for transcript analysis.

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Area of Science:

  • Molecular Biology
  • Biochemistry

Background:

  • mRNA quantitation is crucial for gene transfer studies.
  • mRNA protection assays using end-labeled DNA probes are established methods.
  • In vitro synthesized antisense RNA probes offer an alternative approach.

Purpose of the Study:

  • To highlight the utility of RNA probes for mRNA quantitation and structural analysis.
  • To discuss the advantages of RNA probes over DNA probes in gene expression studies.

Main Methods:

  • Utilizing mRNA protection assays.
  • Employing in vitro transcribed RNA probes.
  • Leveraging bacteriophage RNA polymerases (SP6, T3, T7) for probe synthesis.

Main Results:

  • RNA probes provide a sensitive method for mRNA quantitation.

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  • This technique yields structural information about the target transcript.
  • Bacteriophage RNA polymerases facilitate high-yield, specific labeled transcript synthesis.

    • Conclusions:

    • In vitro synthesized RNA probes are a popular and efficient alternative to DNA probes.
    • The characterization of bacteriophage RNA polymerases has enabled widespread use of RNA probes.
    • These polymerases offer advantages in purification, stability, polymerization rate, and promoter specificity.