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Single Molecule Sequencing Of M13 Virus Genome Without Amplification

By Luyang Zhao, Liwei Deng, Gailing Li, Huan Jin, Jinsen Cai, Huan Shang, Yan Li, Haomin Wu, Weibin Xu, Lidong Zeng, Renli Zhang, Huan Zhao, Ping Wu, Zhiliang Zhou, Jiao Zheng, Pierre Ezanno, Qin Yan, Michael Deem, Jiankui He

Posted 03 May 2017
bioRxiv DOI: 10.1101/133843 (published DOI: 10.1371/journal.pone.0188181)

Third generation sequencing is a direct measurement of DNA/RNA sequences at the single molecule level without amplification. In this study, we report sequencing of the genome of the M13 virus by a new single molecule sequencing platform. Our platform detects single molecule fluorescence by the total internal reflection microscope technique, with sequencing-by-synthesis chemistry. We sequenced the genome of M13 to a depth of 316x and 100% coverage. The consensus sequence accuracy is 100%. We demonstrated that single molecule sequencing has no significant GC bias.

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