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Evaluation of RNA Isolation Methods in Human Adipose Tissue
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences, Clinical diabetology and metabolism.ORCID iD: 0000-0001-8489-5881
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences, Clinical diabetology and metabolism.
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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
Available from: 2022-12-15 Created: 2022-12-15 Last updated: 2025-02-20Bibliographically approved

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Sarsenbayeva, AsselJernow, HenningHetty, SusannePereira, Maria J.

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Clinical diabetology and metabolismDepartment of Medical Sciences
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