Collection and storage of human blood and adipose for genomic analysis of clinical samples

Authors

  • Ann K. Cashion,

    Corresponding author
    1. College of Nursing, University of Tennessee Health Science Center, 920 Madison, Suite 507 N, Memphis, TN 38163
    • College of Nursing, University of Tennessee Health Science Center, 920 Madison, Suite 507 N, Memphis, TN 38163.
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    • Professor.

  • Reba A. Umberger,

    1. College of Graduate Health Sciences, University of Tennessee Health Science Center, Memphis, TN
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    • Doctoral Candidate.

  • Shirlean B. Goodwin,

    1. W. Harry Feinstone Center for Genomic Research, University of Memphis, Memphis, TN
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    • Research Assistant Professor.

  • Thomas R. Sutter

    1. W. Harry Feinstone Center for Genomic Research, University of Memphis, Memphis, TN
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    • Professor.


  • This project was conducted by team members from The University of Tennessee Health Science Center (Deborah Gibson, Robin Bloodworth), Methodist University Hospital Transplant Institute (James Eason), and the University of Memphis W. Harry Feinstone Center for Genomic Research (Quynh Tran).

  • Parts of this manuscript have been presented at the International Society for Nurses in Genetics annual meeting.

Abstract

In this methods article, we describe collection and storage of clinically acquired blood and adipose samples for transcript analysis in an ongoing study exploring obesity in renal transplant recipients. Total ribonucleic acid (RNA) was isolated from whole blood using the LeukoLOCK™ Total RNA Isolation System (n = 4), and comparisons between fresh and frozen samples were made. Abdominal subcutaneous adipose samples (n = 4) were obtained during kidney transplantation, flash frozen, and stored at −80°C. Adipose RNA was extracted using either the STAT-60 method modified for lipids or Trizol plus RNeasy extraction. Affymetrix HG-U133 plus 2.0 arrays and Affymetrix Human Gene 1.0 ST arrays were used for both blood and adipose transcriptome analysis. Purity, quality, and quantity of RNA were high with comparable results using both array platforms. © 2011 Wiley Periodicals, Inc. Res Nurs Health 34:408–418, 2011

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