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Related Experiment Videos

Purification of total RNA from human stool samples

R J Alexander1, R F Raicht

  • 1Department of Veterans Affairs Medical Center, New York, New York 10010, USA.

Digestive Diseases and Sciences
|January 9, 1999
PubMed
Summary
This summary is machine-generated.

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Researchers developed a novel method to isolate RNA from human stool, enabling the analysis of gene expression in shed cells. This technique may offer a less invasive approach for early colon cancer detection.

Area of Science:

  • Molecular Biology
  • Gastroenterology
  • Biochemistry

Background:

  • Colonoscopy is effective for early colon tumor detection but is invasive.
  • A less invasive and more cost-effective method for mucosal sampling is needed.
  • Stool contains sloughed-off colonic cells, potentially carrying tumor-specific gene expression alterations.

Purpose of the Study:

  • To develop a method for isolating total RNA from human stool.
  • To confirm the presence of human gene transcripts in isolated stool RNA.
  • To assess the potential of this method for colon cancer risk assessment.

Main Methods:

  • Developed a stool RNA isolation protocol involving lysis, phenol extraction, and RNA-binding resin.
  • Treated isolated RNA with RNase-free DNase I.

Related Experiment Videos

  • Assayed for human RNA using quantitative slot blotting, northern blotting, and RT-PCR.
  • Validated RNA isolation by detecting rat-specific mRNA in a mixed stool sample.

    • Main Results:

    • Successfully isolated 5-30 microg RNA per gram of stool from cancer patients and 5 microg/g from controls.
    • Quantitative slot blotting indicated approximately 10% of isolated RNA was of human origin.
    • Northern blotting and RT-PCR confirmed the presence of human RNA transcripts.
    • RT-PCR unambiguously detected rat-specific mRNA, validating the isolation method.

    Conclusions:

    • RNA can be successfully isolated from human stool.
    • Human gene transcripts are detectable in stool-derived RNA preparations.
    • This RNA isolation technique offers a promising, non-invasive tool for identifying individuals at risk for colon cancer.