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Emergence of Bias During the Synthesis and Amplification of cDNA for scRNA-seq

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Single Cell Biomedicine

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 1068))

Abstract

The advent of single-cell omics technology has promoted our understanding of the genomic, epigenomic, and transcriptomic heterogeneity in individual cells. Compared to traditional sequencing studies using bulk cells, single-cell transcriptome technology is naturally more dynamic for in depth analysis of genomic variation resulting from cell division and is useful in unraveling the regulatory mechanisms of gene networks in many diseases. However, there are still some limitations of current single-cell RNA sequencing (scRNA-seq) protocols. Biases that arise during the RNA reverse transcription and cDNA pre-amplification steps are the most common problems and play pivotal roles in limiting the quantitative accuracy of scRNA-seq. In this review, we will describe how these biases emerge and impact scRNA-seq protocols. Moreover, we will introduce several current and convenient modified scRNA-seq methods that allow for bias to be decreased and estimated.

Authors Qiankun Luo and Hui Zhang have equally contributed to this chapter as the first author.

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Conflicts of Interest Statement

Qiankun Luo and Hui Zhang have no conflicts of interest.

Author information

Authors and Affiliations

  1. Department of General Surgery, Second Affiliated Hospital, Zhengzhou University, Zhengzhou, China

    Qiankun Luo

  2. Department of Science Research and Discipline Construction, Henan Provincial People’s Hospital, Jinshui District, Zhengzhou, China

    Hui Zhang

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  1. Qiankun Luo
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  2. Hui Zhang
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Editors and Affiliations

  1. Henan Provincial People’s Hospital, Zhengzhou, Henan, China

    Jianqin Gu

  2. Shanghai Institute of Clinical Bioinformatics, Zhongshan Hospital, Fudan University, Shanghai, China

    Xiangdong Wang

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Luo, Q., Zhang, H. (2018). Emergence of Bias During the Synthesis and Amplification of cDNA for scRNA-seq. In: Gu, J., Wang, X. (eds) Single Cell Biomedicine. Advances in Experimental Medicine and Biology, vol 1068. Springer, Singapore. https://doi.org/10.1007/978-981-13-0502-3_12

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