Agarose gel electrophoresis method for detecting microsatellite DNA by using multiple sample application and double-layer electrophoresis

A technology of agarose gel and double-layer electrophoresis, which is applied to measurement devices, material analysis by electromagnetic means, instruments, etc., can solve the problems of waste of agarose powder and electrophoresis buffer, low utilization rate of EB, etc., and achieve rapid detection. , the effect of improving utilization, saving manpower and cost

Inactive Publication Date: 2014-07-23
QIQIHAR UNIVERSITY
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Problems solved by technology

[0005] The purpose of the present invention is to solve the problems of a large amount of waste of agarose powder and electrophoresis buffer and the low utilization rate of EB caused by the existing microsatellite DNA electrophoresis detection, and provide a multi-point sample, double-layer electrophoresis detection microsatellite DNA Agarose Gel Electrophoresis Method

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  • Agarose gel electrophoresis method for detecting microsatellite DNA by using multiple sample application and double-layer electrophoresis
  • Agarose gel electrophoresis method for detecting microsatellite DNA by using multiple sample application and double-layer electrophoresis
  • Agarose gel electrophoresis method for detecting microsatellite DNA by using multiple sample application and double-layer electrophoresis

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specific Embodiment approach 1

[0027] Specific embodiment one: a kind of multi-point sample of the present embodiment, the agarose gel electrophoresis method that double-layer electrophoresis detects microsatellite DNA is carried out according to the following steps:

[0028] 1. Using microsatellite molecular markers to perform PCR amplification on the genomic DNA of different populations or different individuals in the population to obtain the amplified products to be detected;

[0029] 2. Prepare two 3% agarose gels of the same area, take one of the 3% agarose gels as the lower agarose gel, and place the other 3% agarose gel on the lower agarose gel in parallel As the upper layer of agarose gel, put the lower layer of agarose gel and the upper layer of agarose gel into the electrophoresis buffer and set aside;

[0030] 3. After loading the amplified product to be detected in step 1 on the lower agarose gel, perform electrophoresis. When the distance between the amplified product and the spotting hole is 1...

specific Embodiment approach 2

[0040] Embodiment 2: This embodiment differs from Embodiment 1 in that: the electrophoresis voltage in steps 1 to 5 is 7-8 V / cm. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0041] Embodiment 3: This embodiment differs from Embodiment 1 or Embodiment 2 in that the temperature of the electrophoresis buffer described in step 2 is 0-4°C. Others are the same as in the first or second embodiment.

[0042] In order to fully disclose the multi-point sample fast and high-efficiency agarose gel electrophoresis method of the present invention, the present invention is further described in detail by following tests:

[0043] The agarose gel electrophoresis method that a kind of multipoint sample of this test, double-layer electrophoresis detects microsatellite DNA is carried out according to the following steps:

[0044] 1. Extraction of plant DNA samples:

[0045] The all-female thick-skinned melon "WI998" from the United States was used as the female parent, and the dioecious thin-skinned melon "3-2-2" from the West Melon Breeding Laboratory of Northeast Agricultural University was used as the male parent. Methods The muskmelon recombinant inbred line po...

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Abstract

The invention discloses an agarose gel electrophoresis method for detecting microsatellite DNA by using multiple sample application and double-layer electrophoresis, and relates to an agarose gel electrophoresis method for detecting the microsatellite DNA, to solve the problems that agarose powdered sugar and electrophoretic buffer solution are greatly wasted and the EB (Electron Beam) is low in utilization rate during the conventional microsatellite DNA electrophoresis detection. The method comprises the following steps of: putting two pieces of sepharose gel of the same areas on two up and down layers in parallel; carrying out sample application and electrophoresis on the lower layer and the upper layer in sequence till detected amplification products reach the bottom of the gel, stopping the electrophoresis; and subsequently carrying out gel imaging detection. By utilizing the method, the utilization rate of the sepharose gel and the electrophoresis buffer solution is greatly improved, and the labor and the cost are saved; compared with one-time sample application, no gel needs to be reprepared, one piece of 10-cm gel can be repeated for sample application for 8 times, so that the cost can be saved by more than 8 times. Therefore, rapid and efficient detection plant SSR (Solid State Relay) marker-site heritable variation is realized.

Description

technical field [0001] The invention relates to an agarose gel electrophoresis method for detecting microsatellite DNA. Background technique [0002] In many research fields, including genetic linkage map construction, QTL or gene mapping, gene map cloning, phylogenetic evolution, molecular evolution, molecular marker-assisted breeding, genetic diversity analysis, kinship identification, etc., a large number of molecular marker analyzes are involved. This is often something that a laboratory is unable and impossible to undertake independently. Therefore, the development of high-throughput molecular markers is required. High-throughput molecular markers refer to molecular markers that can provide a large amount of genetic information per unit time and processing. Automated analysis is the basis for the development of high-throughput molecular labeling technology. Multi-point sampling is to load multiple polymerase chain reaction products on the same electrophoresis lane to ...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/447
Inventor 高美玲袁成志王世发王芳赵芳芳
Owner QIQIHAR UNIVERSITY
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