Evaluation of RNA Isolation Methods in Human Adipose TissueShow others and affiliations
2022 (English)In: Laboratory medicine, ISSN 0007-5027, E-ISSN 1943-7730, Vol. 53, no 5, p. E129-E133Article in journal (Refereed) Published
Abstract [en]
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 phenolchloroform 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 (similar to 2.0) compared to the traditional method (similar to 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.
Place, publisher, year, edition, pages
Oxford University Press, 2022. Vol. 53, no 5, p. E129-E133
Keywords [en]
RNA isolation, adipose tissue, RNA quantity, RNA purity, traditional, phenol-chloroform method, column-based methods
National Category
Biochemistry Molecular Biology
Identifiers
URN: urn:nbn:se:uu:diva-490822DOI: 10.1093/labmed/lmab126ISI: 000764225900001PubMedID: 35150274OAI: oai:DiVA.org:uu-490822DiVA, id: diva2:1719371
2022-12-152022-12-152025-02-20Bibliographically approved