DNA banking: the effects of storage of blood and isolated DNA on the integrity of DNA.

Author(s): Madisen L, Hoar DI, Holroyd CD, Crisp M, Hodes ME

Publication: Am J Med Genet, 1987, Vol. 27, Page 379-90

PubMed ID: 3605221 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine the effects of storage conditions, temperature and duration on DNA yield and integrity from blood and spleen tissue specimens.

Conclusion of Paper

Generally, storage of whole blood at -70 degrees C did not affect DNA yield or integrity, but when packed cells or whole blood were stored at 4 or 23 degrees C, there was a linear decrease in DNA yield observable at 3 days (first time point), regardless of storage conditions, and the mean DNA yield was lower in blood specimens shipped at ambient temperatures from France to Indianapolis than in the specimens collected from the next 18 patients and processed in France within 24 h. Storage of cell lysates for 2 weeks (shortest duration examined) at 65 degrees C resulted in severe degradation of the DNA, but DNA from lysates stored at 4 or 37 degrees C for 8 weeks remained intact, and storage for an additional 5 days at 25 degrees C did not result in DNA degradation. There was no effect of storing DNA for up to 6 months, over chloroform, at -70, -20, 4, 25 or 37 degrees C. In contrast, DNA stored at 45, 55, or 65 degrees C showed degradation of larger bands after 9 weeks, 6 weeks, and 21 h, respectively.

Studies

  1. Study Purpose

    The purpose of this study was to determine the effects of delayed processing due to transport and storage of whole blood at 3 temperatures, packed cells at 2 temperatures, leukocyte lysates at 3 temperatures, and isolated DNA at 8 different temperatures on the DNA yield and integrity. After experimental storage, some leukocyte lysates were stored for an additional 5 days at 25 degrees C.

    Summary of Findings:

    Generally, storage of whole blood at -70 degrees C for up to 8 weeks did not affect DNA yield or integrity, but 1 of the 6 specimens consistently had a much lower yield (average 2 versus 25 ug/mL) in specimens stored frozen (2, 4, or 8 weeks) than in the corresponding fresh specimen; however, when packed cells or whole blood were stored at 4 or 23 degrees C, there was a linear decrease in DNA yield observable at 3 days (first time point), regardless of storage conditions. Further, the mean DNA yield was lower in 18 blood specimens shipped at ambient temperatures from France to Indianapolis (6 days) than in the specimens collected from the next 18 patients and processed in house within 24 h. Storage of cell lysates for 2 weeks (shortest duration examined) at 65 degrees C resulted in severe degradation of the DNA, but DNA from lysates stored at 4 or 37 degrees C for 8 weeks remained intact, and storage for an additional 5 days at 25 degrees C did not result in DNA degradation. There was no effect of storage of DNA for up to 6 months, over chloroform, at -70, -20, 4, 25 or 37 degrees C. In contrast, DNA that was stored at 45, 55, or 65 degrees C showed degradation of larger bands after 9 weeks, 6 weeks, and 21 h, respectively.

    Biospecimens
    Preservative Types
    • None (Fresh)
    • Other Preservative
    • Frozen
    Diagnoses:
    • Not specified
    Platform:
    AnalyteTechnology Platform
    DNA Electrophoresis
    DNA Southern blot
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Storage Between site transportation method Mailed
    Not transported
    Storage Storage temperature 4 degrees C
    23 degrees C
    25 degrees C
    37 degrees C
    45 degrees C
    55 degrees C
    65 degrees C
    -70 degrees C
    -20 degrees C
    Storage Storage duration 0 h
    21 h
    3 days
    4 days
    5 days
    7 days
    8 days
    2 weeks
    3 weeks
    4 weeks
    5 weeks
    6 weeks
    7 weeks
    8 weeks
    9 weeks
    10 weeks
    26 weeks
    Storage Storage conditions DNA over chloroform
    Leukocyte lysate
    Whole blood
    Packed cells
    Biospecimen Preservation Type of fixation/preservation Frozen
    None (fresh)
    Refrigeration
  2. Study Purpose

    The purpose of this study was to determine the effects of storage of air-dried DNA from spleen tissue for 34 years on DNA integrity. The DNA was resuspended by mixing for 2 weeks at 4 degrees C in a Tris-EDTA solution and the molecular weight of stored DNA was indirectly compared with the authors knowledge of the molecular weight of fresh DNA.

    Summary of Findings:

    The average molecular weight of DNA from a single specimen of splenic DNA stored air-dried for 34 years was <1 kb, but the authors state this may be attributable to mechanical disruption during resuspension.

    Biospecimens
    Preservative Types
    • Other Preservative
    Diagnoses:
    • Not specified
    Platform:
    AnalyteTechnology Platform
    DNA Electrophoresis
    DNA Southern blot
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Storage Storage duration 0 years
    34 years

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