Title: A comparison of microRNA sequencing reproducibility and noise reduction using mirVana and TRIzol isolation methods

Authors: Yan Guo; Amma Bosompem; Xue Zhong; Travis Clark; Yu Shyr; Annette S. Kim

Addresses: Department of Cancer Biology, Vanderbilt University, Nashville, TN 37232, USA ' Department of Pathology, Microbiology and Immunology, Vanderbilt University, Nashville, TN 37232, USA ' Department of Cancer Biology, Vanderbilt University, Nashville, TN 37232, USA ' Vanderbilt Technologies for Advanced Genomics, Vanderbilt University, Nashville, TN 37232, USA ' Department of Biostatistics, Vanderbilt University, Nashville, TN 37232, USA ' Department of Pathology, Microbiology and Immunology, Vanderbilt University, Nashville, TN 37232, USA

Abstract: MicroRNAseq (miRNAseq) is a form of RNAseq technology that has become an increasingly popular alternative to miRNA expression profiling. Unlike messenger RNA (mRNA), miRNA extraction can be difficult, and sequencing such small RNA can also be problematic. We designed a study to test the reproducibility of miRNAseq technology and the performance of the two popular miRNA isolation methods, mirVana and TRIzol, by sequencing replicated samples using microRNA isolated with each kit. Through careful analysis of our data, we found excellent repeatability of miRNAseq technology. The mirVana method performed better than TRIzol in terms of useful reads sequenced, number of miRNA identified, and reproducibility. Finally, we identified a baseline noise level for miRNAseq technology; this baseline noise level can be used as a filter in future miRNAseq studies.

Keywords: next generation sequencing; miRVana isolation; TRIzolv isolation; microRNA sequencing; sequence reproducibility; noise reduction; miRNAseq.

DOI: 10.1504/IJCBDD.2014.061642

International Journal of Computational Biology and Drug Design, 2014 Vol.7 No.2/3, pp.102 - 112

Published online: 27 May 2014 *

Full-text access for editors Access for subscribers Purchase this article Comment on this article