A Simple Approach for Evaluating Total MicroRNA Extraction from Mouse Brain Tissues

Jack G. Walleshauser III, Trace Kessler, Danielle Morse, Bakhos A. Tannous, Norman H. L. Chiu

Department of Chemistry and Biochemistry, University of North Carolina at Greensboro, Greensboro, USA.
Department of Nanoscience, Joint School of Nanoscience and Nanoengineering, Greensboro, USA.
Molecular Neuro- genetics Unit, Department of Neurology, Massachusetts General Hospital and Harvard Medical School, Boston, USA.

DOI: 10.4236/jasmi.2012.21002                

Abstract

For the analysis of microRNA, a common approach is to first extract microRNA from cellular samples prior to any specific microRNA detection. Thus, it is important to determine the quality and yield of extracted microRNA. In this study, solid-phase extraction was used to isolate small RNA (<200 nt), which included microRNA, from mouse brain tissues. By using standard UV absorbance measurements, the amount of small RNA in the extracted RNA samples was determined. To determine the presence of microRNA, each RNA sample was analyzed by PAGE with SYBR^? Green II staining. Testing for contamination of any small DNA fragments, RNase and cellular peptides or proteins were systematically carried out. By scanning the gel image obtained from PAGE analysis, the average percentage of total microRNA (19 - 25 nt) in the extracted RNA samples was determined to be equal to 2.3 ± 0.5%. The yield of total microRNA was calculated to be ~0.5ng of microRNA per milligram of frozen mouse brain tissue. In comparison to other methods that require the use of expensive specialized instrumentation, the approach of combining the standard UV absorbance and PAGE analysis represents a simple and viable method for evaluating the quality and yield of microRNA extraction from tissue samples.

Keywords

MicroRNA; Extraction; Tissue

Conflicts of Interest

The authors declare no conflicts of interest.

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