Evaluation of RNA Isolation Methods in Human Adipose Tissue

Abstract

Objective: Research has shown that RNA extraction from adipose tissue (AT) is challenging because of high lipid content and low RNA quantity. We compared a traditional RNA extraction with a column-based method in human AT to evaluate RNA quantity and quality.

Materials and methods: Human subcutaneous AT (n = 9) was collected through needle biopsy, and RNA was extracted using the phenol-chloroform traditional method and the RNeasy Lipid Tissue Mini Kit column-based method. The RNA quantity, quality, integrity, and expression of key AT genes were assessed.

Results: We found that the RNA quantity and integrity were reduced by 40% and 15-20%, respectively, using the column-based method compared to the traditional method, but the findings were not statistically significant. The column-based method showed a higher 260/280 ratio (~2.0) compared to the traditional method (~1.8) (P <.05), suggesting lower amounts of contaminants. The expression of AT genes was comparable between methods.

Conclusion: The traditional extraction method provides adequate RNA yield and integrity compared to the column-based method, which is an advantage when AT specimens are small.

Keywords: RNA isolation; RNA purity; RNA quantity; adipose tissue; column-based methods; traditional phenol-chloroform method.