College of Nursing, University of Tennessee Health Science Center, 920 Madison, Suite 507 N, Memphis, TN 38163, USA.
Res Nurs Health. 2011 Oct;34(5):408-18. doi: 10.1002/nur.20448. Epub 2011 Aug 2.
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.
在本方法学文章中，我们描述了一项正在进行的研究中临床采集的血液和脂肪样本的收集和储存，该研究旨在探索肾移植受者中的肥胖问题。使用 LeukoLOCK™ 总 RNA 分离系统从全血中分离总核糖核酸 (RNA)（n=4），并比较了新鲜样本和冷冻样本之间的差异。在肾移植过程中获得腹部皮下脂肪样本（n=4），立即冷冻并储存在-80°C。使用改良的 STAT-60 法（用于脂质）或 Trizol 加 RNeasy 提取法从脂肪组织中提取 RNA。使用 Affymetrix HG-U133 plus 2.0 阵列和 Affymetrix Human Gene 1.0 ST 阵列分别对血液和脂肪转录组进行分析。两种阵列平台的 RNA 纯度、质量和数量均较高，结果具有可比性。
