Preparation method of genome mixing sequencing library

A technology for sequencing libraries and genomes, applied in the field of high-throughput sequencing technology, can solve the problems of exceeding the construction cost of a single sample library and the low amount of data in a single reaction, so as to improve the connection efficiency, improve the efficiency of library construction, and save the cost of library construction. Effect

Inactive Publication Date: 2016-06-15
WUHAN BINGGANG BIOTECH CO LTD
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Problems solved by technology

The characteristic of 454 sequencing is that the length of a single read grows, but the amount of data in a single reaction is low
However, for some small genomes, such as fungal genomes with a size of about 2.5-81.15Mb and organelle genomes such as chloroplast genome

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  • Preparation method of genome mixing sequencing library
  • Preparation method of genome mixing sequencing library
  • Preparation method of genome mixing sequencing library

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Embodiment Construction

[0044] Below in conjunction with accompanying drawing and embodiment of description, specific embodiment of the present invention is described in further detail:

[0045] Below in conjunction with specific embodiment, further elaborate the present method invention. These examples are only used to illustrate the present invention and are not intended to limit the scope of protection of the present invention. The specific experimental conditions and methods are not indicated in the following examples, and the experimental conditions and methods refer to relevant reagent instructions or "Molecular Cloning Experiment Guide" (3rd edition) (J. Sambrook, edited by D.W. Russell, published in 2008).

[0046] Implementation of an example of preparation of a mixed-sample sequencing library for four samples of Solanaceae Dilania (DC) samples.

[0047] Table 2. Sample, tag sequence and index sequence information table in mixed sample library preparation

[0048]

[0049] The four samp...

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Abstract

The invention relates to the technical field of molecular biology, and discloses a method for preparing a genome mixed-sample sequencing library, which comprises the following steps: (1) ultrasonic fragmentation of genomic DNA; (2) purification and end repair of fragmented products; (3) ) repair product for purification and adapter ligation; (4) ligation product purification recovery and concentration determination; (5) sample mixing and fragment screening; (6) PCR amplification of the product after fragment screening; (7) purification of the PCR product to obtain Sequencing library; (8) library quality inspection and on-machine sequencing. The present invention provides an adapter compatible with the Illumina next-generation sequencer in the above step (2), a method for purifying the product in the step (2) (3) (4) (7), and a method for mixing samples in the step (4) And the PCR amplification system and program setting in the step (6) ensure that the library building method provided by the present invention can quickly and smoothly carry out multiple sample mixed sample library building, and the method provided by the present invention can be used to obtain good data uniformity. and high-quality sequencing data.

Description

technical field [0001] The invention relates to the field of high-throughput sequencing technology in molecular biology. More specifically, it relates to a method for preparing a genome mixed sample sequencing library, which is suitable for resequencing or simplified sequencing of various samples of all eukaryotic organisms, especially for resequencing of small genes with multiple samples or simplified sequencing of large gene groups. Background technique [0002] Next-generation sequencing technology is currently the most commonly used technology in high-throughput sequencing research. After more than 30 years of development, DNA sequencing technology has made significant progress, and the second-generation sequencing technology characterized by high throughput has gradually matured and become commercialized. Early sequencing technologies mainly relied on first-generation sequencing. The various DNA sequencing technologies developed from the traditional chemical degradatio...

Claims

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Application Information

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IPC IPC(8): C40B50/06C12Q1/68
CPCC40B50/06C12Q1/6869
Inventor 张洪源郑媛坤束礼平杨冰范艺翔王宇峰何银竹束礼伟黄刚董扬
Owner WUHAN BINGGANG BIOTECH CO LTD
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