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Rapid genotype identification method suitable for map-based cloning

A map-based cloning and identification method technology, applied in the field of genetics, can solve problems such as time-consuming and duplication of genotype identification methods, and achieve the effects of accelerating work progress, reducing duplication of labor, and saving cost and time

Active Publication Date: 2020-02-11
MAIZE RES INST SHANDONG ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] For this reason, the technical problem to be solved by the present invention is to provide a method for rapid extraction of genomic DNA and rapid identification of genotype suitable for map-based cloning, to solve the problem of repetitive and time-consuming methods for extraction of genomic DNA and identification of genotype in the prior art. question

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  • Rapid genotype identification method suitable for map-based cloning
  • Rapid genotype identification method suitable for map-based cloning
  • Rapid genotype identification method suitable for map-based cloning

Examples

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Effect test

Embodiment 1

[0033] The genotype rapid identification method suitable for map-based cloning described in this embodiment comprises the following steps:

[0034] (1) Boil the towel in boiling water for 15 minutes, remove it after cooling thoroughly, and spread half of it on the bottom of a plastic box with a length of about 25 cm and a width of about 18 cm; Then spread the seeds (about 500 or so) evenly on the towel; the other half of the towel is covered on the seeds. Place the plastic box in a light incubator for cultivation (25°C, 16 hours of light, 8 hours of darkness);

[0035] (2) Sampling was taken at 7-10 days after germination, and each corn seedling was taken to a length of about 1cm above the growth point and put into a 2ml centrifuge tube (numbered in advance; stainless steel beads were installed, diameter 6mm), and each centrifuge tube put 10 Corn seedling samples (numbered 1-10 in sequence); get a repetition, and set aside later;

[0036] (3) Break the sample with a tissue d...

Embodiment 2

[0053] The genotype rapid identification method suitable for map-based cloning described in this embodiment comprises the following steps:

[0054] (1) Boil the towel in boiling water for 15 minutes, remove it after cooling thoroughly, and spread half of it on the bottom of a plastic box about 25cm long and 18cm wide; the shrunken endosperm grains are first soaked with ultra-pure water 2-3 times, Then spread the seeds (about 500 or so) evenly on the towel; the other half of the towel is covered on the seeds. Place the plastic box in a light incubator for cultivation (25°C, 16 hours of light, 8 hours of darkness);

[0055] (2) Sampling was taken at 7-10 days after germination, and each corn seedling was taken to a length of about 1cm above the growth point and put into a 2ml centrifuge tube (numbered in advance; stainless steel beads were installed, diameter 6mm), and each centrifuge tube put 10 Corn seedling samples (numbered 1-10 in sequence); get a repetition, and set aside...

Embodiment 3

[0073] The genotype rapid identification method suitable for map-based cloning described in this embodiment comprises the following steps:

[0074] (1) Boil the towel in boiling water for 15 minutes, remove it after cooling thoroughly, and spread half of it on the bottom of a plastic box about 25cm long and 18cm wide; the shrunken endosperm grains are first soaked with ultra-pure water 2-3 times, Then spread the seeds (about 500 or so) evenly on the towel; the other half of the towel is covered on the seeds. Place the plastic box in a light incubator for cultivation (25°C, 16 hours of light, 8 hours of darkness);

[0075] (2) Sampling was taken at 7-10 days after germination, and each corn seedling was taken to a length of about 1cm above the growth point and put into a 2ml centrifuge tube (numbered in advance; stainless steel beads were installed, diameter 6mm), and each centrifuge tube put 10 Corn seedling samples (numbered 1-10 in sequence); get a repetition, and set aside...

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Abstract

The invention belongs to the technical field of genetics and specifically relates to a rapid genome DNA extraction and genotype identification method suitable for map-based cloning. The rapid genotypeidentification method suitable for map-based cloning utilizes crop seedlings as verification samples, performs verification with a general DNA extraction method and performs PCR amplification throughreasonably designed primers, so that the method can rapidly finish the rapid extraction of genome DNA and the rapid identification of genotypes. Thus, repetitive work can be greatly reduced, costs and time can be saved, work progress can be accelerated; and the method can be used for reference in other crops as well.

Description

technical field [0001] The invention belongs to the technical field of genetics, and in particular relates to a method for rapid extraction of genomic DNA and rapid genotype identification suitable for map-based cloning. Background technique [0002] The acquisition of the draft genome of maize in February 2008 made maize the second crop successfully sequenced after rice. In 2009, American scientists completed the genome sequencing of maize inbred line B73, and its coverage reached 95% (Schnable et al.2009). It is predicted that the size of the maize genome is about 2,500MB, and the number of genes is about 50,000-60,000. The sequencing team has already stored the draft information of the maize genome in the Internet gene sequencing public database maizeGDB, and is constantly updating the data. [0003] The large-scale sequencing of the maize genome is conducive to the development of enough molecular markers by using the genome sequence information. At the same time, the ge...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/686C12Q1/6806
CPCC12Q1/6806C12Q1/686C12Q2565/125C12Q2527/125
Inventor 关海英汪黎明刘铁山何春梅刘春晓董瑞刘强王娟
Owner MAIZE RES INST SHANDONG ACAD OF AGRI SCI
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