RNA isolation from adipose tissue: An optimized procedure for high RNA yield and integrity

Abstract

Adipose tissue physiology strongly differs between body regions. Reasonable amounts of intact RNA are required to analyze the local differences on a molecular level. Therefore, we developed an optimized RNA isolation procedure from adipose tissue samples. Excised human subcutaneous adipose tissue was obtained from elective operations, and the RNA Stabilization Reagent (RNAlater) was tested for its effect on RNA integrity. Additionally, 3 different RNA isolation kits were evaluated for efficacy in isolating RNA from tissue samples. The samples displayed a significant loss in recoverable RNA and signs of RNA degradation 30 to 60 minutes after tissue excision. The application of RNAlater significantly delayed this degradation. Using the RNeasy Lipid Tissue Kit resulted in a significantly higher RNA yield compared to the RNeasy Mini Kit. Our results demonstrate that combining the RNAlater and RNeasy Lipid Tissue Kit results in a higher yield of RNA even from relatively small tissue samples with guaranteed RNA integrity.