Journal article
Identification of an optimal method for extracting RNA from human skin biopsy, using domestic pig as a model system
Scientific Reports, Vol.9(1)
2019
Abstract
To evaluate skin tissue gene expression patterns correctly, extracting sufficient quantities of good quality RNA is essential. However, RNA extraction from skin tissue is challenging, as the hyaluronic acid-collagen matrix is extremely difficult to homogenize. Although there are multiple ways to extract RNA from skin, there are no comparative studies that identify the most critical steps, e.g. sample collection, storage and homogenization. We analysed the various steps involved in RNA extraction (i.e. biopsy collection as dry biopsy or into nucleotide stabilizing reagents, different storage conditions, enzymatic digestion, stator-rotor and bead motion-based homogenizing combined with column-based RNA purification). We hypothesised that domestic pig skin is applicable as a model for human skin studies. Altogether twenty different workflows were tested on pig skin and the four most promising workflows were tested on human skin samples. The optimal strategy for extracting human skin RNA was to collect, store and homogenize the sample in RLT lysis buffer from the RNeasy Fibrous Tissue Kit combined with beta-mercaptoethanol. Both stator-rotor and bead motion-based homogenizing were found to result in high quality and quantity of extracted RNA. Our results confirmed that domestic pig skin can be successfully used as a model for human skin RNA studies.
Details
- Title
- Identification of an optimal method for extracting RNA from human skin biopsy, using domestic pig as a model system
- Authors/Creators
- E. Reimann (Author/Creator) - University of TartuK. Abram (Author/Creator) - Tartu University HospitalS. Kõks (Author/Creator) - Murdoch UniversityK. Kingo (Author/Creator) - Tartu University HospitalA. Fazeli (Author/Creator) - University of Sheffield
- Publication Details
- Scientific Reports, Vol.9(1)
- Publisher
- Springer Nature
- Identifiers
- 991005539985707891
- Copyright
- © 2020 Springer Nature Limited
- Murdoch Affiliation
- Centre for Comparative Genomics
- Language
- English
- Resource Type
- Journal article
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- Collaboration types
- Domestic collaboration
- International collaboration
- Citation topics
- 1 Clinical & Life Sciences
- 1.54 Molecular & Cell Biology - Genetics
- 1.54.1454 PCR Techniques
- Web Of Science research areas
- Dermatology
- ESI research areas
- Clinical Medicine