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Method for conducting enrichment library construction and SNP analysis on gene of complex genome of higher plant

A higher plant and genome technology, applied in biochemical equipment and methods, organic compound libraries, recombinant DNA technology, etc.

Active Publication Date: 2015-02-18
BGI TECH SOLUTIONS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When studying these complex genomes, the genomic library obtained by the conventional RAD library construction method contains a large number of repetitive sequences, which cannot play the role of enriching the genomic gene region
[0005] Although there are already methods for enriching higher plant genome gene regions in the prior art, such as MF (Methylated Filtration) (Palmer et al., 2003) for filtering out methylated sequences, and HC (High-Cot) for filtering out high-copy sequences (Yuan et al., 2003), HMPR (Hypomethylated Partial Restriction), etc., but the results of Emberton et al. (2005) showed that the library gene sequences constructed by the above three methods can only reach 33.8% and 23.4% of the total sequence respectively. %, 25.8%

Method used

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  • Method for conducting enrichment library construction and SNP analysis on gene of complex genome of higher plant
  • Method for conducting enrichment library construction and SNP analysis on gene of complex genome of higher plant
  • Method for conducting enrichment library construction and SNP analysis on gene of complex genome of higher plant

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Experimental program
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Effect test

Embodiment

[0043] Materials: Four varieties of corn are included, namely B73, Nongkebai, Zheng 58, and 7922. Plant one plant of each variety.

[0044] See Table 1, Table 2, Table 3, and Table 4 for the reagents and instruments used in this example.

[0045] Table 1

[0046]

[0047]

[0048] All the above reagents were stored at -20°C.

[0049] Table 2

[0050]

[0051] The above reagents were stored at room temperature.

[0052] table 3

[0053]

[0054] All the above reagents were stored at 4°C.

[0055] Table 4 List of commonly used instruments

[0056]

[0057] Proceed as follows:

[0058] 1. Take the leaves of maize when it grows for one month, and extract maize DNA with the CTAB extraction method proposed by Doyle JJ and Doyle JL (1987). The steps are briefly described as follows: 1) Take 2×CTAB and β-mercaptoethanol 25:1 (volume) in a centrifuge tube and preheat; 2) Put 1-2 g of corn leaves into a mortar precooled by liquid nitrogen, Add liquid nitrogen and...

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Abstract

The invention provides a method of library construction suitable for gene regions of a higher flowering plant containing a complex genome. In the method of the invention, the gene regions of the genome are enriched by a methylation-sensitive enzyme instead of a conventional non-methylation-sensitive enzyme, and then a library is constructed. The invention also provides a method, suitable for a higher flowering plant containing a complex genome, for searching SNP.

Description

technical field [0001] The invention belongs to the field of molecular biology, in particular to a method for establishing a complex genome gene library of higher plants. Background technique [0002] Restriction-site associated DNA (RAD) labeling technology refers to a technique for sequence analysis of the restriction site region after genomic DNA is digested with restriction endonucleases. In the early stage of RAD marker application, gene chip technology was used to separate RAD markers. For example, Lewis et al. (2007) used NotI to digest the DNA of N. crassa, then used a gene chip to perform sequence analysis on the NotI restriction site region, and completed the mapping of the mutation site; Miller et al. ( 2007) analyzed the EcoRI markers of the zebrafish genome with gene chip technology, and also completed the mapping of mutation sites; Miller et al. (2007) tested the genotyping of biological monomers and isolated populations using RAD gene chips, and the results ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C40B50/06C40B40/06C12N15/10C12Q1/68
CPCC12N15/1034C12N15/1093
Inventor 郭钰邵迪韩长磊陶晔杨焕明张秀清
Owner BGI TECH SOLUTIONS