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White flower beet the 9th chromosome BAC chip and its preparation method

A technology of chromosome number and chromosome, which is applied in the field of BAC chip of white flower beet chromosome 9 and its preparation, and can solve the problems of density limitation of membrane chip, incomparability of signal value, etc.

Inactive Publication Date: 2005-01-26
HEILONGJIANG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

On the other hand, the density limit of the membrane chip requires the same probe to hybridize dozens of membranes, which leads to incomparability of signal values ​​between different membranes.

Method used

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Experimental program
Comparison scheme
Effect test

specific Embodiment approach 1

[0004] Specific embodiment 1: In this embodiment, the BAC clone on the BAC chip of chromosome 9 of the beet is to use the scattered repetitive sequence of the chromosome of the beet as the specific probe to screen the BAC DNA obtained from the BAC library. The aforementioned BAC DNA is the same as the carrier. It is obtained by compounding. In this embodiment, the 2377 BAC DNA obtained by screening the BAC library by using the scattered repetitive sequences of the chromosomes of white beet as a specific probe are all compounded with the carrier. The carrier is a solid phase carrier. The solid phase of this embodiment The carrier is a glass slide. Its preparation steps are: (1) BAC preliminary treatment: a. Dissolve 5-10μg of purified and extracted BAC DNA in 50μl of 0.1-0.15M NaOH solution, and in a water bath at 60-70℃ for 30-60 minutes B. Add 5μl of 0.1-0.2M glycidoxytrimethoxysilane dissolved in isopropanol; c. Water bath in a 60-70℃ water bath for 3-5 hours; e. Add 50μl of neu...

specific Embodiment approach 2

[0009] Specific implementation manner 2: Using the scattered repetitive sequences of the chromosomes of the beet as a specific probe to screen the BAC library, a part of the 2377 BAC DNAs are compounded with the vector.

specific Embodiment approach 3

[0010] Specific embodiment 3: The carrier in this embodiment is a silicon wafer, a nylon membrane, a nitrocellulose membrane or a gel.

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Abstract

A white bloom beet chromosome 9 BAC chip and its preparing method are provided. BAC clone on the BAC chip is obtained by screening the BAC library using white bloom beet chromosome, and the chip is prepared by combining the BAC clone with the carrier. The preparing method comprises: primary treatment for BAC, BAC treatment before spotting, and preparation of the BAC chip. M14 and M14 chromosome addition line differential expression gene screening, genome differential analysis, gene mapping and number of replications analysis can be done rapidly and efficiently by using the chip prepared by the method.

Description

Technical field: [0001] The present invention relates to the field of biotechnology, and in particular to a BAC chip of chromosome 9 of beet beet and a preparation method thereof. Background technique: [0002] Plant apomixis (Apomixis) refers to a special asexual reproduction method that reproduces by seeds without fusion of sperm and eggs. Since there is no diploid offspring produced by the fusion of sperm and egg, their genotype is exactly the same as that of the mother, so that the offspring inherit the inherent genetic characteristics of the parent and form a genetically stable clone that can be reproduced by seeds [2]. Plant apomixis because of its special asexual reproduction through seeds, it can fix favorable genotypes, change breeding procedures and seed production, which has great economic significance. If it can be applied to important crops, it will be a successor to the sixties of the twentieth century. After the "Green Revolution" in the 1980s, another agricultural...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 顾雪锋王志伟李海英韩俊郭德栋朱大海
Owner HEILONGJIANG UNIV
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