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
Advanced SearchBrowse by AnalyteBrowse by Pre-analytical FactorSearch SOPsSOP Compendiums

Biological stability of mRNA isolated from human postmortem brain collections.

Author(s): Leonard S, Logel J, Luthman D, Casanova M, Kirch D, Freedman R

Publication: Biol Psychiatry, 1993, Vol. 33, Page 456

PubMed ID: 8098224 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine if RNA extracted from postmortem brain specimens varying in postmortem interval, delay prior to freezing, and frozen storage duration can be successfully used in reverse transcription-polymerase chain reaction (RT-PCR), Northern blot, and in vitro translation analyses.

Conclusion of Paper

The authors report that RNA extracted from brains stored for approximately 5 years at -70 degrees C may exhibit some degree of degradation but is of sufficient quality to perform adequately in polymerase chain reaction (PCR) amplification. They further report that frozen storage and/or prolonged dissection times may lead to degradation too extensive for RNA use in in vitro expression or oligo-dT primed library construction.

Studies

  1. Study Purpose

    The purpose of this study was to evaluate the influence of postmortem interval, on-ice dissection time, and frozen storage duration on the quality and quantity of RNA isolated from postmortem brain tissue.

    Summary of Findings:

    The authors report that postmortem interval, dissection time delay, and frozen storage time did not influence RNA yield. The amount of degradation detected via Northern blot for beta-actin mRNA increased with the duration of frozen storage but not postmortem interval or dissection time. The authors recommend that postmortem brain specimens used for in vitro translation should be collected within 1 hour of death and subjected to frozen storage for no more than 1 year, although optimal results were obtained with freshly collected specimens. PCR amplification of APRT, bFGF, and FGFr genes in random hexamer primered samples was successful among all specimen handling groups.

    Biospecimens
    Preservative Types
    • Frozen
    Diagnoses:
    • Autopsy
    Platform:
    AnalyteTechnology Platform
    RNA Northern blot
    RNA RT-PCR
    RNA Spectrophotometry
    RNA First-strand cDNA Synthesis
    RNA In vitro translation
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Preaquisition Postmortem interval 7 h
    14 h
    21 h
    30.5 h
    40.5 h
    42.5 h
    Storage Storage duration > 3 months at -70 degrees C
    4 - 6 years at -70 degrees C
    Storage Storage duration Dissection time delay of 1.25 h
    Dissection time delay of 3.75 h
    Dissection time delay of 5.00 h
    Dissection time delay of 6.00 h
    View more

You Recently Viewed  

News and Announcements

  • Most Downloaded SOPs in 2024
  • New Articles on the GTEx Project are Now FREELY Available!
  • Just Published!
    • More...