Successful RNA extraction from various human postmortem tissues.

Author(s): Heinrich M, Matt K, Lutz-Bonengel S, Schmidt U

Publication: Int J Leg Med, 2007, Vol. 121, Page 136

PubMed ID: 17115174 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to optimize RNA extraction from postmortem skeletal muscle, brain, heart, liver, kidney, and spleen.

   Summary of Findings:

   In optimizing the RNA extraction protocol, the authors found it necessary to treat the extract with DNase to remove genomic DNA; they also found useful a clean-up step with RNeasy prior to reverse transcription.

   Biospecimens

   • Tissue - Muscle (Skeletal)
   • Tissue - Spleen
   • Tissue - Liver
   • Tissue - Kidney
   • Tissue - Brain
   • Tissue - Heart

   Preservative Types

   • Frozen

   Diagnoses:

   • Autopsy
   • Not specified

   Platform:

   Analyte	Technology Platform
   RNA	Electrophoresis

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Analyte Extraction and Purification	Analyte isolation method	With RNeasy clean-up Without RNeasy clean-up
   Analyte Extraction and Purification	Nucleic acid digestion	DNase treatment No DNase treatment

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2. Study Purpose

   The purpose of this study was to evaluate whether an association could be made between postmortem interval and RNA yield and integrity in postmortem skeletal muscle, brain, heart, liver, kidney, and spleen.

   Summary of Findings:

   The authors report successful amplification of GAPDH transcripts from most tissue types from each postmortem interval studied. No correlation was observed between postmortem interval and RNA degradation. Highest RNA yields were obtained from muscle, brain, and liver. RNA degradation, determined by successful RT-PCR amplification of a range of product sizes, was independent of postmortem interval, although some tissue-specific results were observed. Generation of a GAPDH 453 bp amplicon was not always possible in spleen, while brain and muscle yielded 619 bp amplicons in all instances.

   Biospecimens

   • Tissue - Muscle (Skeletal)
   • Tissue - Spleen
   • Tissue - Liver
   • Tissue - Kidney
   • Tissue - Brain
   • Tissue - Heart

   Preservative Types

   • Frozen

   Diagnoses:

   • Autopsy
   • Not specified

   Platform:

   Analyte	Technology Platform
   RNA	RT-PCR
   RNA	Real-time qRT-PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Acquisition	Biospecimen location	Skeletal Muscle Brain Heart Liver Kidney Spleen
   Preaquisition	Postmortem interval	15 h 22 h 26 h 36 h 56 h 118 h
   RT-PCR Specific	Length of gene fragment	114 bp 233 bp 453 bp 619 bp 866 bp

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