Weipeng Mo
Junpeng Shi
Ning Song
Pei Liang
Jian Chen
Yiting Shi
Weilong Guo
Xinchen Li
Xiaohong Yang
Beibei Xin
Haiming Zhao
Weibin Song
Jinsheng Lai
a State Key Laboratory of Plant Physiology and Biochemistry and National Maize Improvement Center, Department of Plant Genetics and Breeding, China Agricultural University, Beijing 100193, PR China;
b Department of Microbiology and Immunology, College of Biological Sciences, China Agricultural University, Beijing 100193, PR China;
c State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing 100193, PR China;
d Key Laboratory of Crop Heterosis and Utilization, State Key Laboratory for Agrobiotechnology, Beijing Key Laboratory of Crop Genetic Improvement, China Agricultural University, Beijing 100193, PR China;
e Center for Crop Functional Genomics and Molecular Breeding, China Agricultural University, Beijing 100193, PR China
Funds: This work was supported by grants from National Key Research and Development Program (2016YFD0101803 - 04) and National Natural Science Foundation of China (31421005 and 91935303).
Received Date: 2021-04-12
Accepted Date:2021-05-13
Rev Recd Date:2021-05-03
Publish Date:2021-08-20
Abstract
Abstract
DNA sequencing is vital for many aspects of biological research and diagnostics. Despite the development of second and third generation sequencing technologies, Sanger sequencing has long been the only choice when required to precisely track each sequenced plasmids or DNA fragments. Here, we report a complete set of novel barcoding and assembling system, Highly-parallel Indexed Tagmentation-reads Assembled Consensus sequencing (HITAC-seq), that could massively sequence and track the identities of each individual sequencing sample. With the cost of much less than that of single read of Sanger sequencing, HITAC-seq can generate high-quality contiguous sequences of up to 10 kilobases or longer. The capability of HITAC-seq was confirmed through large-scale sequencing of thousands of plasmid clones and hundreds of amplicon fragments using approximately 100 pg of input DNAs. Due to its long synthetic length, HITAC-seq was effective in detecting relatively large structural variations, as demonstrated by the identification of a ～1.3 kb Copia retrotransposon insertion in the upstream of a likely maize domestication gene. Besides being a practical alternative to traditional Sanger sequencing, HITAC-seq is suitable for many high-throughput sequencing and genotyping applications.Keywords: HITAC-seq,
Structure variation,
Sequencing technology,
Sanger sequencing,
Comparative genomics
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