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Analysis of the Molecular Quality of Human Tissues. An Experience From the Cooperative Human Tissue Network

Author(s): Jewell Scott D, Srinivasan Mythily, McCart Linda M, Williams Nita, Grizzle William H, LiVolsi Virginia, MacLennan Greg, Sedmak Daniel D

Publication: Am J Clin Pathol, 2002, Vol. 118, Page 733

PubMed ID: 12428794 PubMed Review Paper? No

Purpose of Paper

To assess nucleic acid quality after prolonged frozen storage (banked tissue), to compare quality determining platforms, and to assess nucleic acid stability after a delay at room temperature prior to processing.

Conclusion of Paper

Authors conclude that gel electrophoresis is a comparable determinant of nucleic acid integrity in comparison to PCR and Northern blot analyses. DNA and RNA stability was not affected by a 5 h delay at room temperature post excision and prior to freezing.

Studies

  1. Study Purpose

    To assess DNA integrity and usability after prolonged frozen storage (0-3, 3-6, 6-12, >12 months) and to compare the efficacy of technical platforms. Tissue types examined included breast, colon, liver, lung, ovary, endometrium, and cervix.

    Summary of Findings:

    DNA integrity, as determined by visual confirmation of a DNA ladder of ~12 kb, was adequate in 79.5% of specimens examined by gel electrophoresis. DNA usability, determined by PCR generation of a 379 bp genomic product (HRPT) was adequate in 78.1% of examined specimens.

    Biospecimens
    Preservative Types
    • Frozen
    Diagnoses:
    • Not specified
    Platform:
    AnalyteTechnology Platform
    DNA PCR
    DNA Electrophoresis
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Storage Storage duration 0-1 month
    3-6 months
    6-12 months
    >12 months
    View more
  2. Study Purpose

    To assess nucleic acid stability after delays at room temperature (15 min, 30 min, and 1, 2, 3, 4, 5 h) after excision and prior to freezing.

    Summary of Findings:

    Even after 5 h at room temperature prior to freezing, DNA and RNA integrity and stability did not differ from controls.

    Biospecimens
    Preservative Types
    • Frozen
    Diagnoses:
    • Not specified
    Platform:
    AnalyteTechnology Platform
    RNA RT-PCR
    DNA PCR
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Storage Time at room temperature 30 min
    1 h
    2 h
    3 h
    4 h
    5 h
    15 min
    View more
  3. Study Purpose

    To assess RNA quality and usability after prolonged frozen storage (0-3, 3-6, 6-12, >12 months) and to compare the efficacy of technical platforms. Tissue types examined included breast, colon, liver, lung, ovary, endometrium, and cervix.

    Summary of Findings:

    RNA quality, as determined by visual confirmation of 28S and 18S ribosomal bands, was adequate in 56.3% of specimens examined. HRPT RT-PCR yielded a 177bp product in 63.6% of tissue examined. Northern blot analysis for G3PDH exhibited a clear and predicted 1.3kb product in 54.3% of examined tissues.

    Biospecimens
    Preservative Types
    • Frozen
    Diagnoses:
    • Not specified
    Platform:
    AnalyteTechnology Platform
    RNA Electrophoresis
    RNA Northern blot
    RNA RT-PCR
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Storage Storage duration 0-3 months
    3-6 months
    6-9 months
    9-12 months
    > 12 months
    View more

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