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Quantitative comparison of single-cell sequencing methods using hippocampal neurons

Luwen Ning, Guan Wang, Zhoufang Li, Wen Hu, Qingming Hou, Yin Tong, Meng Zhang, Li Qin, Xiaoping Chen, Hengye Man, Pinghua Liu, Jiankui He
doi: https://doi.org/10.1101/004366
Luwen Ning
1Department of Biology, South University of Science and Technology of China, Shenzhen 518055, China
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Guan Wang
2Departments of Biology, Boston University, Boston, MA 02215, USA
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Zhoufang Li
1Department of Biology, South University of Science and Technology of China, Shenzhen 518055, China
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Wen Hu
3Departments of Chemistry, Boston University, Boston, MA 02215, USA
4State Key Laboratory of Respiratory Disease, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China
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Qingming Hou
2Departments of Biology, Boston University, Boston, MA 02215, USA
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Yin Tong
1Department of Biology, South University of Science and Technology of China, Shenzhen 518055, China
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Meng Zhang
1Department of Biology, South University of Science and Technology of China, Shenzhen 518055, China
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Li Qin
4State Key Laboratory of Respiratory Disease, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China
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Xiaoping Chen
4State Key Laboratory of Respiratory Disease, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China
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Hengye Man
2Departments of Biology, Boston University, Boston, MA 02215, USA
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  • For correspondence: he.jk@sustc.edu.cn pinghua@bu.edu hman@bu.edu
Pinghua Liu
3Departments of Chemistry, Boston University, Boston, MA 02215, USA
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  • For correspondence: he.jk@sustc.edu.cn pinghua@bu.edu hman@bu.edu
Jiankui He
1Department of Biology, South University of Science and Technology of China, Shenzhen 518055, China
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  • For correspondence: he.jk@sustc.edu.cn pinghua@bu.edu hman@bu.edu
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Abstract

Single-cell genomic analysis has grown rapidly in recent years and will find widespread applications in various fields of biology, including cancer biology, development, immunology, pre-implantation genetic diagnosis, and neurobiology. In this study, we amplified genomic DNA from individual hippocampal neurons using one of three single-cell DNA amplification methods (multiple annealing and looping-based amplification cycles (MALBAC), multiple displacement amplification (MDA), and GenomePlex whole genome amplification (WGA4)). We then systematically evaluated the genome coverage, GC-bias, reproducibility, and copy number variations among individual neurons. Our results showed that single-cell genome sequencing results obtained from the MALBAC and WGA4 methods are highly reproducible and have a high success rate. Chromosome-level and subchromosomal-level copy number variations among individual neurons can be detected.

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Posted April 21, 2014.
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Quantitative comparison of single-cell sequencing methods using hippocampal neurons
Luwen Ning, Guan Wang, Zhoufang Li, Wen Hu, Qingming Hou, Yin Tong, Meng Zhang, Li Qin, Xiaoping Chen, Hengye Man, Pinghua Liu, Jiankui He
bioRxiv 004366; doi: https://doi.org/10.1101/004366
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Quantitative comparison of single-cell sequencing methods using hippocampal neurons
Luwen Ning, Guan Wang, Zhoufang Li, Wen Hu, Qingming Hou, Yin Tong, Meng Zhang, Li Qin, Xiaoping Chen, Hengye Man, Pinghua Liu, Jiankui He
bioRxiv 004366; doi: https://doi.org/10.1101/004366

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