Method for cutting wheat chromosomes by SLmuCUT (molecular machine industry, MMI) system

A chromosome and wheat technology, applied in the field of molecular cytogenetics, can solve the problems of large variation in chromosome structure, huge workload of screening library, low protein content, etc., and achieve the effects of uniform dispersion of chromosomes, rapid production, and simple production.

Inactive Publication Date: 2012-09-19
HENAN UNIV OF SCI & TECH
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Problems solved by technology

[0003] (1) The existence of plant cell walls and the difficulty of synchronization hinder the preparation of well-dispersed chromosome specimens, and the preparation of chromosome specimens is one of the most basic techniques affecting chromosome cutting; the background is not clean, which affects the cutting efficiency
[0004] (2) The ploidy of the plant genome is diverse, the chromosome structure varies greatly, and the sequence of genes is not conservative enough; the DNA content of plant chromosomes is high and the protein content is low, and most chromosomes cannot display G, R or Q bands like vertebrate chromosomes. Affects DNA seque

Method used

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  • Method for cutting wheat chromosomes by SLmuCUT (molecular machine industry, MMI) system
  • Method for cutting wheat chromosomes by SLmuCUT (molecular machine industry, MMI) system
  • Method for cutting wheat chromosomes by SLmuCUT (molecular machine industry, MMI) system

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[0040] 1. Preparation of chromosome specimens of root tip cells

[0041] (1) Rooting: soak the seeds of wheat Yannong 15, China Spring, Xiaoyan Mai Shannong 0095, and Elytrigia intermedium at room temperature until white, freeze in a refrigerator at 1-4°C for 24h, and then cultivate at 25°C. Take the root tip when it is 1.5-2.0cm long; or water the material grown in the field in the evening in warm weather, and dig the root tip from 9-10 in the morning;

[0042] (2) Fixation and storage: the root tip is treated in ice water for 24 hours, then fixed with Carnot's solution for more than 24 hours at 1-4°C, then transferred to 70% alcohol for storage;

[0043] (3) Pretreatment: Take the root tip cells out of the preservation solution and treat them with sterile distilled water for 10 minutes;

[0044] (4) Preliminary microscopic examination: Perform preliminary microscopic examination of the cell division phase of the root tip, and select most of the root tip cells in the middle stage for...

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Abstract

The invention relates to a method for cutting wheat chromosomes by a SLmuCUT (molecular machine industry, MMI) system, comprising the steps of taking root tip cell of common wheat (Yannong 15 and Chinese spring), Trititrigia alien substitution line Shannong 0095, and Thinopyrum intermedium as materials; performing microdissection for wheat root tip cell chromosomes on the basis of using the SLmuCUT system to primarily cut the wheat root tip cell chromosomes. The method is simple, quick and convenient to produce slice, fast in slice production speed, and uniform in chromosomes dispersion; on the basis of using the SLmuCUT system to primarily cut the wheat root tip cell chromosomes, main technical parameters for the cutting system for cutting the wheat root tip cell chromosomes is optimized; the microdissection for wheat chromosomes segment, single chromosome, two/three chromosomes is performed successfully; and therefore, the reference is provided for subsequent research such as microdissection and microcloning of specific wheat chromosomes and wheat non-homologous chromosomes, and cloning of exogenous chromosome carrying excellent gene.

Description

technical field [0001] The invention relates to the establishment of a technical system for cutting wheat chromosomes by using the SLμCUT system, which belongs to the field of molecular cytogenetics. Background technique [0002] Chromosome micro-dissection and micro-cloning (chromosome micro-dissection and micro-cloning) technology was proposed by Scalenghe et al. Melanogaster polytene chromosome [J]. Chromosoma, 1981, 82: 205-216) is a new technology of molecular cytogenetics. This technique first cuts the Drosophila chromosome, and successfully obtained 80 clones, which were then used in mice and humans (Rohme Dan, Howard Fox, Bernhard Herrmann, et al. Molecular clones of the mouse complex derived from microdissected metaphase chromosomes[J ]. Cell, 1984, 36:783-788.). With the development of PCR technology, microdissection and microcloning techniques have been greatly developed (Ludecke HJ, Gabriele Senger, Use Claussen, et al. Cloning defined regions of human genome b...

Claims

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Application Information

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IPC IPC(8): C40B50/06C40B40/06C12Q1/68
Inventor 王黎明李兴锋袁瑛袁建国高居荣王洪刚
Owner HENAN UNIV OF SCI & TECH
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