Feasibility of Whole RNA Sequencing from Single-Cell mRNA AmplificationReport as inadecuate




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Genetics Research InternationalVolume 2013 2013, Article ID 724124, 8 pages

Research Article

Department of Computer Science, MCG, Augusta, GA 30912, USA

Renji Hospital of Shanghai, Jiaotong University School of Medicine, Shanghai, China

School of Computer Engineering, Nanyang Technological University, Singapore 639798

Department of Pediatrics, MCG, Augusta, GA 30912, USA

Received 8 August 2013; Revised 17 October 2013; Accepted 13 November 2013

Academic Editor: Bernard Weissman

Copyright © 2013 Yunbo Xu et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Single-cell sampling with RNA-seq analysis plays an important role in reference laboratory; cytogenomic diagnosis for specimens on glass-slides or rare cells in circulating blood for tumor and genetic diseases; measurement of sensitivity and specificity in tumor-tissue genomic analysis with mixed-cells; mechanism analysis of differentiation and proliferation of cancer stem cell for academic purpose. Our single- cell RNA-seq technique shows that fragments were 250–450 bp after fragmentation, amplification, and adapter addition. There were 11.6 million reads mapped in raw sequencing reads 19.6 million. The numbers of mapped genes, mapped transcripts, and mapped exons were 31,332, 41,210, and 85,786, respectively. All QC results demonstrated that RNA-seq techniques could be used for single-cell genomic performance. Analysis of the mapped genes showed that the number of genes mapped by RNA-seq 6767 genes was much higher than that of differential display 288 libraries among similar specimens which we have developed and published. The single-cell RNA-seq can detect gene splicing using different subtype TGF-beta analysis. The results from using Q-rtPCR tests demonstrated that sensitivity is 76% and specificity is 55% from single-cell RNA-seq technique with some gene expression missing 2-8 genes. However, it will be feasible to use RNA-seq techniques to contribute to genomic medicine at single-cell level.





Author: Yunbo Xu, Hongliang Hu, Jie Zheng, and Biaoru Li

Source: https://www.hindawi.com/



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