Murdoch University Research Repository

Welcome to the Murdoch University Research Repository

The Murdoch University Research Repository is an open access digital collection of research
created by Murdoch University staff, researchers and postgraduate students.

Learn more

Identification of an optimal method for extracting RNA from human skin biopsy, using domestic pig as a model system

Reimann, E., Abram, K., Kõks, S., Kingo, K. and Fazeli, A. (2019) Identification of an optimal method for extracting RNA from human skin biopsy, using domestic pig as a model system. Scientific Reports, 9 (1).

[img]
Preview
PDF - Published Version
Download (1MB) | Preview
Free to read: https://doi.org/10.1038/s41598-019-56579-5
*No subscription required

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.

Item Type: Journal Article
Murdoch Affiliation: Centre for Comparative Genomics
Publisher: Springer Nature
Copyright: © 2020 Springer Nature Limited
United Nations SDGs: Goal 9: Industry, Innovation and Infrastructure
URI: http://researchrepository.murdoch.edu.au/id/eprint/54168
Item Control Page Item Control Page

Downloads

Downloads per month over past year