Evaluation of RNA Isolation Methods in Human Adipose Tissue.

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.