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Yield and Integrity of RNA from Brain Samples are Largely Unaffected by Pre-analytical Procedures

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Abstract

Gene expression studies are reported to be influenced by pre-analytical factors that can compromise RNA yield and integrity, which in turn may confound the experimental findings. Here we investigate the impact of four pre-analytical factors on brain-derived RNA: time-before-collection, tissue specimen size, tissue collection method, and RNA isolation method. We report no significant differences in RNA yield or integrity between 20 mg and 60 mg tissue samples collected in either liquid nitrogen or the RNAlater stabilizing solution. Isolation of RNA employing the TRIzol reagent resulted in a higher yield compared to isolation via the QIAcube kit while the latter resulted in RNA of slightly better integrity. Keeping brain tissue samples at room temperature for up to 160 min prior to collection and isolation of RNA resulted in no significant difference in yield or integrity. Our findings have significant practical and financial consequences for clinical genomic departments and other laboratory settings performing large-scale routine RNA expression analysis of brain samples.

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Acknowledgements

Emil Jakobsen is greatly acknowledged for his support euthanizing the mice and excision of their brains. The authors also thank the staff at the department of Genomic Medicine at Copenhagen University Hospital for their support and guidance.

Funding

This work was in part funded by the Novo Nordisk Foundation (NNF14CC0001), Department of Drug design and Pharmacology, Copenhagen University and the Center of Genomic Medicine, Rigshospitalet, Copenhagen, Denmark. The funders had no role in study design, data collection and analysis, decision to publish or preparation of manuscript.

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  1. Pernille Søs Hovgaard Jensen and Maja Johansen have been contributed equally to this publication.

Authors and Affiliations

  1. Department of Technology, Faculty of Health and Technology, University College Copenhagen, 2200, Copenhagen, Denmark

    Pernille Søs Hovgaard Jensen, Maja Johansen & Christina Kjær

  2. Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, 2100, Copenhagen, Denmark

    Lasse K. Bak & Christina Kjær

  3. Disease Systems Biology Program, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, 2200, Copenhagen, Denmark

    Lars Juhl Jensen

Authors
  1. Pernille Søs Hovgaard Jensen
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  3. Lasse K. Bak
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  4. Lars Juhl Jensen
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  5. Christina Kjær
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Contributions

MJ cut the tissue and PSHJ collected it. PSHJ and MJ performed QIAcube and TRIzol isolation of RNA, measurements of RNA yield and integrity and attributed to results. LKB analyzed the data. CK, LKB and LJJ interpreted the results. CK did the conceptual work. CK and LJJ designed the experiments, with contribution from PSHJ and MJ. CK wrote the draft of the paper. All authors provided input on the manuscript and approved the final version.

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Correspondence to Christina Kjær.

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Jensen, P.S.H., Johansen, M., Bak, L.K. et al. Yield and Integrity of RNA from Brain Samples are Largely Unaffected by Pre-analytical Procedures. Neurochem Res 46, 447–454 (2021). https://doi.org/10.1007/s11064-020-03183-z

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  • DOI: https://doi.org/10.1007/s11064-020-03183-z

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