NIH, National Cancer Institute, Division of Cancer Treatment and Diagnosis (DCTD) NIH - National Institutes of Health National Cancer Institute DCTD - Division of Cancer Treatment and Diagnosis
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Recovery and expression of messenger RNA from postmortem human brain tissue.

Author(s): Cummings TJ, Strum JC, Yoon LW, Szymanski MH, Hulette CM

Publication: Mod Pathol, 2001, Vol. 14, Page 1157-61

PubMed ID: 11706078 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine if the quantity and quality of RNA extracted from brain specimens is affected by postmortem interval prior to refrigeration, patient diagnosis, cerebrospinal pH, or premortem oxygen administration.

Conclusion of Paper

The authors report that all 19 specimens evaluated yielded intact RNA that did not display significant degradation, regardless of premortem factors such as Alzheimer's dementia, fever, cerebrospinal pH, oxygen administration, prolonged illness, sepsis, or postmortem interval prior to refrigeration (range 1 to 14 hours). Although the quantity of extracted RNA and edg-1 gene expression exhibited a high degree of inter-specimen variability, results were not correlated to postmortem interval, cerebrospinal pH or any of the premortem factors examined.

Studies

  1. Study Purpose

    The purpose of this paper was to determine if the quantity and quality of RNA extracted from brain specimens is affected by postmortem interval prior to refrigeration, patient diagnosis, cerebrospinal pH, or premortem oxygen administration. Gene expression levels of a representative immediate early gene, edg-1, was also investigated.

    Summary of Findings:

    RNA degradation was not observed among any specimens. While the quantity of total RNA, as determined by spectrophotometer, exhibited a high degree of variation among specimens (as large as an order of magnitude), no correlation to postmortem interval prior to refrigeration, cerebrospinal fluid pH, or premortem oxygen administration was observed. Real time quantitative RT-PCR analysis of edg-1 expression resulted in high specimen variability (by as much as 8-fold), although no clear correlation was observed among any of the factors examined. Of note, RNA used as template for real time qRT-PCR was quantified using the RNA specific dye Ribogreen.

    Biospecimens
    Preservative Types
    • Frozen
    Diagnoses:
    • Neoplastic
    • Not specified
    • Alzheimer's Disease
    • Autopsy
    Platform:
    AnalyteTechnology Platform
    RNA Spectrophotometry
    RNA Electrophoresis
    RNA Real-time qRT-PCR
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Preaquisition Diagnosis/ patient condition Alzheimer's disease
    No dementia
    Fever
    Sepsis
    Other illnesses
    Preaquisition Postmortem interval 1 h 10 min - 3 h 19 min
    7 h 25 min - 10 h 48 min
    12 h 16 min - 14 h 00 min
    Biospecimen Aliquots and Components pH 5
    6
    7
    8
    Preaquisition Other drugs Supplemental oxygen
    No supplemental oxygen
    View more

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