Building method of DNA large-fragment library for Pacbio platform and assay kit

A library construction and large fragment technology, which is applied to the construction of large DNA fragment libraries. The kit field of this library construction method can solve the problems of high cost of library construction and sequencing, and limited application range, so as to improve the purity and reduce the cost of library construction. Cost, flexible and convenient effect
CN107794575AInactive Publication Date: 2018-03-13BGI GENOMICS CO LTD
4 Cites 9 Cited by

Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
BGI GENOMICS CO LTD
Publication Date
2018-03-13
Estimated Expiration
Not applicable · inactive patent

Smart Images

  • Figure 1
    Figure 1
  • Figure 2
    Figure 2
  • Figure 3
    Figure 3
Patent Text Reader

Abstract

The application discloses a building method of DNA large-fragment library for Pacbio platform and an assay kit. The building method includes subjecting broken DNA large fragments to VII enzyme treatment, sequentially carrying out primary DNA damage repairing and terminal repairing treatment, linker splicing treatment, exonuclease ExoIII and ExoVII double-digest treatment, subjecting products obtained by double-digest treatment to fragment separation through nucleic acid electrophoresis and a fragment recovery system, subjecting products obtained by the fragment separation to secondary DNA damage repairing, so as to obtain the DNA large-fragment library adaptable to the Pacbio platform. According to the building method of the DNA large-fragment library, the whole building process is carriedout without Pacbio library-building assay kit, library building cost is lowered greatly, and a material foundation is laid for promotion and application of the third-generation sequencing and Pacbionucleic acid sequencing platform.
Need to check novelty before this filing date? Find Prior Art

Description

technical field

[0001] This application relates to the field of nucleic acid sequencing, in particular to a method for constructing a DNA large fragment library for the Pacbio platform, and a kit especially for the method for constructing a library. Background technique

[0002] The Pacbio platform, that is, the Pacbio nucleic acid sequencing platform, uses the third-generation sequencing technology, that is, single molecule real-time sequencing (Single Molecule Real-Time). The sequencing of the Pacbio platform is based on two key technologies. First, a nanopore (zero-modewaveguides, abbreviated as ZMW) with a diameter of only tens of nanometers invented by Pacific Biosciences, in which only single molecule nucleic acid can be allowed to pass through the DNA. The reaction is carried out under the action of polymerase, and the four fluorescently labeled deoxynucleotides of A, T, C, and G in the synthesis process in the nanopore are detected to realize nucleic acid sequencing;...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More