Resequencing the Escherichia coli genome by GenoCare single molecule sequencing platform


Luyang Zhao, Liwei Deng, Gailing Li, Huan Jin, Jinsen Cai, Huan Shang, Yan Li, Andrew X. Yang, Fang Chen, Zhi Zhao1, Guanjie Xu, Wuxing Liu, Siyu Liu, Guobing Xiang, Bin Liu, Weibin Xu, Lidong Zeng, Renli Zhang, Huan Zhao, Ping Wu, Zhiliang Zhou, Jiao Zheng, Pierre Ezanno, Weiyue Chen, Qin Yan, Michael W. Deem, Jun Yu5, Jiankui He

 

Next generation sequencing (NGS) has revolutionized life sciences research. Recently, a new class of third-generation sequencing platforms has arrived to meet increasing demands in the clinic, capable of directly measuring DNA and RNA sequences at the single-molecule level without amplification. Here, we use the new GenoCare single molecule sequencing platform from Direct Genomics to resequence the E. coli genome and show comparable performance to the Illumina MiSeq system. Our platform detects single-molecule fluorescence by total internal reflection microscopy, with sequencing-by-synthesis chemistry. With a consensus sequence of 99.71% nucleotide identity to that of the Illumina MiSeq system’s, GenoCare was determined to be a reliable platform for single-molecule sequencing, with strong potential for clinical applications. Single-Molecule DNA Sequencing of a Viral Genome.