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

Res Nurs Health. 2011 Oct;34(5):408-18. doi: 10.1002/nur.20448. Epub 2011 Aug 2.

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.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Adipose Tissue / metabolism*
  • Blood Preservation / methods*
  • Blood Specimen Collection / methods*
  • Freezing
  • Gene Expression Profiling / methods*
  • Humans
  • RNA / isolation & purification*
  • Sensitivity and Specificity
  • Tissue Array Analysis / methods

Substances

  • RNA