Stability of RNA isolated from human trabecular bone at post-mortem and surgery.

Author(s): Kuliwaba JS, Fazzalari NL, Findlay DM

Publication: Biochim Biophys Acta, 2005, Vol. 1740, Page 1

PubMed ID: 15878735 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to asses the influence of postmortem interval on RNA quality.

   Summary of Findings:

   The authors report successful isolation of RNA from all specimens, which had postmortem intervals up to 84 hours. Tailing of the RNA on electrophoretic gels, indicative of degradation, did not correlate with patient age, postmortem interval or storage duration. When one 24-hour-postmortem case was stored at 4 degrees C for up to 96 hours, RT-PCR evaluation of RANKL, OPG, IL-6, TGF-beta1, and GAPDH, with the exception of IL-6, indicated declining RNA levels over time, albeit at differing rates. Reverse-transcriptase polymerase chain reaction (RT-PCR), was successfully performed despite some RNA degradation, which was not the case for Northern blot analysis.

   Biospecimens

   • Tissue - Bone

   Preservative Types

   • None (Fresh)

   Diagnoses:

   • Autopsy

   Platform:

   Analyte	Technology Platform
   RNA	Northern blot
   RNA	Electrophoresis
   RNA	RT-PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Preaquisition	Postmortem interval	4 - 84 hours 24 h 48 h 72 h 96 h

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2. Study Purpose

   The purpose of this study was to evaluate the influence of differing bone storage conditions on extracted RNA for bone specimens obtained surgically.

   Summary of Findings:

   The authors report that a 4 degrees C storage temperature is preferable to room temperature or 37 degrees C. Northern blot and RT-PCR analysis of RANKL, OPG, RANK, CTR, OCN, IL-6, TGF-beta1, and GAPDH indicated that, in general, these RNA species degraded by 48 hours at 4 degrees C and more rapidly at warmer temperatures.

   Biospecimens

   • Tissue - Bone

   Preservative Types

   • None (Fresh)

   Diagnoses:

   • Osteoarthritis
   • Osteoporosis

   Platform:

   Analyte	Technology Platform
   RNA	Northern blot
   RNA	Electrophoresis
   RNA	RT-PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Storage temperature	4 degrees C Room temperature 37 degrees C
   Storage	Storage duration	2 h 4 h 8 h 16 h 24 h 48 h 72 h

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