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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Northern and Southern blot analysis of human RNA and DNA in autopsy material.

Author(s): Larsen S, Rygaard K, Asnaes S, Spang-Thomsen M

Publication: APMIS, 1992, Vol. 100, Page 498

PubMed ID: 1376998 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine if RNA and DNA obtained from seven postmortem tissues with differing postmortem intervals (PMI; 21-118 h) can be successfully analyzed by Northern blot analysis for retinoblastoma and beta-actin gene expression, and Southern blot analysis for the retinoblastoma gene.

Conclusion of Paper

Northern blot analysis was successful for all tissues with a PMI of 2 days or less, and was dependent on the degree of autolysis and the presence of two distinct ribosomal bands. Southern blot analysis was independent of autolysis, and was PMI- and tissue- dependent: all tissues with a PMI of less that 75 h resulted in successful Southern blot analysis, while for a PMI of 5 days only lung, adrenal gland, and brain yielded results. Brain samples alone appeared independent of postmortem interval, yielding Northern and Southern blot results in almost all of the specimens evaluated.

Studies

  1. Study Purpose

    The purpose of this paper was to determine if RNA and DNA obtained from seven postmortem tissues (heart, brain, lung, liver, spleen, kidney, adrenal gland) with differing postmortem intervals (PMI; 21-118 h) can be successfully analyzed by Northern blot analysis for retinoblastoma and beta-actin gene expression, and Southern blot analysis for the retinoblastoma gene.

    Summary of Findings:

    Ribosomal RNA bands were successfully visualized using gel electrophoresis for most specimens within 50 hours postmortem. Northern blot probes for retinoblastoma and beta-actin genes were successful for specimens which displayed two distinct ribosomal RNA bands. Histological preservation was a good predictor of successful RNA evaluation. Southern blots for retinoblastoma DNA was successful for all specimens up to 75 hours postmortem, and up to 5 days postmortem for brain, lung, and adrenal glands. The success of these assays did not correlate with histological preservation.

    Biospecimens
    Preservative Types
    • Frozen
    Diagnoses:
    • Autopsy
    • Not specified
    Platform:
    AnalyteTechnology Platform
    RNA Northern blot
    Morphology Light microscopy
    RNA Electrophoresis
    DNA Southern blot
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Acquisition Biospecimen location Brain
    Heart
    Lung
    Kidney
    Spleen
    Liver
    Adrenal gland
    Preaquisition Postmortem interval 21h
    24 h
    26 h
    33 h
    35 h
    38 h
    50 h
    74 h
    118 h
    View more

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