High throughput oligonucleotide sequencing method

一种寡核苷酸、高通量的技术，应用在生物化学设备和方法、微生物的测定/检验等方向，能够解决费用大、局限、制约SAGE作用等问题

Active Publication Date: 2009-05-20
SHENGZHEN CHINA GENE TECH COMPANY
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Problems solved by technology

These various ligase-based DNA sequencing methods can be used to perform parallel sequencing of monoclonal DNA arrays constructed by the methods described above, however, the cost of sequencing remains a huge barrier to the application of sequencing technology at the genome level
[0007] The problem with the existing technology is that while oligonucleotide arrays have contributed enormously to advances in functional genomics and systems biology, there are fundamental limitations to this technology
The cost is also very high, which seriously restricts the role of SAGE, and it is almost impossible to expand its application to clinical medicine.

Method used

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example 1

[0077] In order to identify the template, the beads must be transferred to a sequencing system. After the PCR picoliter reaction chamber hood is directly removed from the horizontal surface, the PCR method or the highly enriched microbead array on the flat surface is fixedly attached, and the sample is sealed in the flow chamber. Use an ITO glass slide to press against the flow cell.

[0078] In an exemplary system, we developed a method based on amine chemistry for the one-step immobilization of DNA-bearing microbeads on a specially modified glass surface. This approach works very well, with a fill factor approaching 70% of the available surface, and excess beads can be used without overlapping issues. What's more, unbound beads are easily recovered for future experiments without losing their functionality. The bead-covered slide is placed in the sequencing flow chamber, which seals itself by simple surface adhesion. This differs from those methods that rely on pressure se...

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Abstract

The present invention relates to the field of gene engineering, provides a high throughout oligonucleotides sequencing method. Said sequencing method includes following steps: the generation of a short DNA tag, single-molecule PCR amplification benefit-marking sequence, enrichment of high-density DNA tags and large-scale cycle parallel sequencing, etc. By using a molecular scale for parallel sequencing in a sequencing method which takes complexation as the base or expanded synthesis as the base, the scale or an expanded anchor primer is used for expanding the base points which are read out on a template. By using the primer, longer sequence on the template can be read out, and the only identification can be obtained from the genome sequence of various organisms with the range from microorganisms, plants, and animals to human, thereby having greater practical value.

Description

field of invention [0001] The invention relates to a gene sequencing method, in particular to a high-throughput oligonucleotide sequencing method. Background technique [0002] As the number of genomes that have been sequenced increases, it is necessary to analyze a large amount of information encoded by genome sequences and the biological functions controlled and participated by these different sequence elements scattered in the genome. However, this is more complex than genome sequencing and requires powerful tools and systematic approaches that can handle processes with a large number of parameters and under different conditions, both normal and abnormal. Only such a comprehensive research method can make us understand most of the basic principles of biology. Of course, such research will also lead to major technological advances for the benefit of mankind. [0003] A major method developed in functional genomics research involves the quantitative analysis of DNA sequen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6874C12Q2525/191C12Q2525/179C12Q2521/501C12Q2565/531
Inventor 邵志峰盛司潼
Owner SHENGZHEN CHINA GENE TECH COMPANY
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