Comparative analysis of single-cell parallel sequencing approaches in oocyte application. (February 2019)

• Record Type: Journal Article
• Authors: Qian, Yan; Liao, Jinyue; Suen, Ashley Hoi Ching; Lee, Annie Wing Tung; Chung, Hoi Sze; Tang, Nelson Leung Sang; Chow, King Lau; Cao, Qin; Yip, Yuk Lap; Leung, Tak Yeung; Chan, Wai-Yee; Chan, David Yiu Leung; Li, Tin Chiu; Lee, Tin-Lap
• Is Part Of: International journal of biochemistry & cell biology. Volume 107(2019)
• Page Start: 1
• Page End: 5
• Publication Date: 2019-02
• Subjects: Single cell parallel sequencing -- Oocyte -- RNA-seq -- Reduced representation bisulfite sequencing -- Whole genome bisulfite sequencing; Biochemistry -- Periodicals; Cytology -- Periodicals; Biochemistry -- Periodicals; Cell Biology -- Periodicals; Biochimie -- Périodiques; Cytologie -- Périodiques; Biochimie; Cytologie; Biochemistry; Cytology; Ressource Internet (Descripteur de forme); Périodique électronique (Descripteur de forme); Periodicals; 572.05
• DOI: 10.1016/j.biocel.2018.12.003
• Languages: English
• ISSNs: 1357-2725

Abstract

Graphical abstract: Abstract: Single-cell parallel sequencing allows us to explore how genetic and epigenetic variations correlate of gene expression in the same cell. Beads-based approach and non-beads-based approach are the two present methods to separate DNA and RNA from the same cell. However, systematic difference between the two methods are lacking. In our study, we compared the performances of the two methods using transcriptome and methylome profiles generated simultaneously from single mouse oocytes. Our results showed that the beads-based approach could capture maximum quantity of mRNA but loss of DNA was inevitable, while the non-beads-based approach could obtain more DNA due to the undamaged nucleus obtained but at a cost of partial loss of mRNA. As the sequencing coverage of methylome sequencing in a single cell was relatively low, single-cell whole genome bisulfite sequencing (scWGBS) was preferable to generate the methylome map in single-cell parallel sequencing in comparison to single-cell reduced representation bisulfite sequencing (scRRBS). To the best of our knowledge, this is the first study to compare the two methods of single-cell parallel sequencing which offers a basic idea for deciding between the two methods and a direction of single-cell parallel sequencing development.