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RNA Quality Control Using External Standard RNA.

Takema Hasegawa1, Junko Takahashi2, Hitoshi Iwahashi1

  • 1Faculty of Applied Biological Sciences, Gifu University, Yanagido, Gifu, Japan.

Polish Journal of Microbiology
|November 20, 2018
PubMed
Summary
This summary is machine-generated.

This study introduces a novel RNA quality control method using external standard RNA to assess messenger RNA (mRNA) yield, enzyme inhibition, and degradation. This approach offers a more accurate evaluation of mRNA quality than traditional methods.

Keywords:
RNA degradationRNA quality controlStandard RNA

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • RNA Analysis

Background:

  • Current RNA quality control methods like RNA Integrity Number and UV absorption do not always accurately reflect messenger RNA (mRNA) quality.
  • Standard RNA, designed based on mRNA, presents an opportunity for improved mRNA quality assessment.

Purpose of the Study:

  • To develop a new RNA quality control method utilizing external standard RNA.
  • To evaluate mRNA yield, DNA polymerase inhibition, and mRNA degradation as key indicators of RNA quality.

Main Methods:

  • Incorporated external standard RNA before the RNA extraction process.
  • Assessed mRNA yield and enzyme inhibition by quantifying standard RNA loss.
  • Evaluated RNA degradation by comparing the 5' and 3' regions of the RNA.

Main Results:

  • Demonstrated that the proposed method can determine mRNA yield and enzyme inhibition.
  • Observed higher degradation at the 3' end compared to the 5' end in Saccharomyces cerevisiae crude extracts.
  • Established that the ratio of degradation between the 3' and 5' ends can quantify the degree of RNA degradation.

Conclusions:

  • External standard RNA provides a viable tool for comprehensive RNA quality control.
  • The new method offers a more accurate assessment of mRNA quality by considering yield, inhibition, and degradation.
  • This approach enhances the reliability of RNA samples for downstream applications.