Stability of messenger RNA in postmortem human brains and construction of human brain cDNA libraries.

Author(s): Kobayashi H, Sakimura K, Kuwano R, Sato S, Ikuta F, Takahashi Y, Miyatake T, Tsuji S

Publication: J Mol Neurosi, 1990, Vol. 2, Page 29

PubMed ID: 1701658 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to evaluate the quantity and quality of poly(A)(+)-RNA in tissue up to 12 hours postmortem.

   Summary of Findings:

   Total RNA and poly (A)(+) RNA yields were not affected significantly by PMI (up to 12 h). Northern blot analysis of neuron, astrocyte, and oligodendrocyte specific genes demonstrated that no reduction in transcript size occurred as a result of PMI. The presence of full length transcripts was confirmed via Southern blot analysis of cDNA libraries. While only one case represesented each preservation temperature, no differences in RNA quality or the successful construction of a cDNA library was observed among specimens frozen in liquid nitrogen or in a -70 degree C freezer.

   Biospecimens

   • Tissue - Brain

   Preservative Types

   • Frozen

   Diagnoses:

   • Pneumonia/Respiratory Infection
   • Autopsy
   • Neoplastic - Leukemia

   Platform:

   Analyte	Technology Platform
   RNA	Northern blot
   RNA	Spectrophotometry
   DNA	Southern blot

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Preaquisition	Postmortem interval	5 h 7 h 9 h 12 h
   Biospecimen Preservation	Cooling or freezing method/ rate	Snap freezing in liquid nitrogen Slow freezing in a -70 degrees C freezer

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