Impact of commercial RNA extraction methods on the recovery of human RNA sequence data from archival fixed tissues.

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Tác giả: Melanie Kuch, Susan Morgello, Debi Poinar, Hendrik Poinar, Jasmine Rockarts, Bruce Wainman, Tess Wilson

Ngôn ngữ: eng

Ký hiệu phân loại: 739.2 Work in precious metals

Thông tin xuất bản: England : BioTechniques , 2025

Mô tả vật lý:

Bộ sưu tập: NCBI

ID: 698454

Archival fixed tissues hold key insights into the evolutionary history of RNA viruses and the associated host immune response, yet access to the RNA sequence data is limited by a lack of robust methods for RNA extraction and sequence retrieval from these tissue types. Here we compared three commercial RNA extraction techniques (bead, column, and phase-based) on five fixed human brain tissues done in triplicate, that have been stored for up to 43 years. We found that for this sample set, bead-based extractions captured longer molecules and yielded a greater proportion of unique reads when aligned to the human genome, than did column and phase-based extraction methods. Via the incorporation of multiple extraction replicates, we quantified the variability in sequencing metrics resulting from tissue sample and extraction technique heterogeneity. Additionally, we compared pre- and post-sequencing metrics and found that the former poorly predicted post-sequencing on-target success. Our findings help inform future research on the recovery of RNA from archival fixed tissues.
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