Elsevier

Analytical Biochemistry

Volume 484, 1 September 2015, Pages 1-3
Analytical Biochemistry

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Purification of high-quality RNA from synthetic polyethylene glycol-based hydrogels

https://doi.org/10.1016/j.ab.2015.05.002Get rights and content

Abstract

Polyethylene glycol (PEG)-based hydrogels, with variable stiffness, are widely used in tissue engineering to investigate substrate stiffness effects on cell properties. Transcriptome analysis is a critical method for understanding cell physiology. However, significant RNA degradation was observed during the process of isolating and purifying RNA from cells encapsulated in the PEG hydrogel, thereby precluding purification of high-quality RNA. Here, we describe a simple protocol that prevents RNA degradation and improves the quality and yield of RNA isolated from cells cultured in PEG hydrogels. This modification produces high-quality total RNA suitable for RNA sequencing and microarray analysis.

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Acknowledgments

This study was supported by National Institute of General Medical Sciences (NIGMS) Grant 1P20GM109091-01 and National Institutes of Health (NIH) grant P30 GM103336 (M.S.), National Science Foundation (NSF) Grant CBET1403545 and NIH Grant AR063745 (E.J.), and a University of South Carolina ASPIRE pilot award (H.J., E.J., and M.S.).

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