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

Authors :
Thomas R. Sutter
Reba Umberger
Shirlean Goodwin
Ann K. Cashion
Source :
Research in Nursing & Health. 34:408-418
Publication Year :
2011
Publisher :
Wiley, 2011.

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

Details

ISSN :
01606891
Volume :
34
Database :
OpenAIRE
Journal :
Research in Nursing & Health
Accession number :
edsair.doi...........7bdb3bb1dcd44dd333a94af5174b4d69
Full Text :
https://doi.org/10.1002/nur.20448