InDel molecular marker primer of Modilong white gourds and application of InDel molecular marker primer

A molecular marker, Modilong technology, applied in the direction of recombinant DNA technology, microbial measurement/testing, biochemical equipment and methods, etc., can solve the problems of prone to errors, poor repeatability, less development and utilization of wax gourd molecular markers, etc.

Active Publication Date: 2020-10-20
HUNAN VEGETABLE RES INST
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Molecular marker methods commonly used in plant genetic research include restriction fragment length polymorphism (RFLP), random amplified polymorphism (RAPD) and simple repeat sequence marker (SSR), but the above-mentioned techniques have the following defects: RFLP requires The participation of radioactive isotopes, the technology is more complicated; the repeatability of RAPD experiment is poor, and the reliability of the result is low; compared with the former two, SSR is simpler, cheaper, and has better repeatability, but some amplification bands are unclear and false, etc. Technical defects such as alleles and null alleles lead to errors in genotyping and data analysis of SSR markers
[0004]

Method used

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  • InDel molecular marker primer of Modilong white gourds and application of InDel molecular marker primer
  • InDel molecular marker primer of Modilong white gourds and application of InDel molecular marker primer
  • InDel molecular marker primer of Modilong white gourds and application of InDel molecular marker primer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Obtaining of InDel molecular markers

[0027] Through the whole genome resequencing of Modilong wax gourd, 466 InDel markers were obtained, 26 of which were selected for PCR amplification, and the primers corresponding to 7 InDel markers obtained electrophoresis bands that were significantly different from those of the parents. , as shown in Table 1 below.

[0028] Table 1 Band characteristics of PCR amplification products corresponding to 7 pairs of InDel marker primers

[0029]

[0030]

[0031] Although the remaining InDel markers can also obtain bands, because the difference between the parents and the offspring is not obvious, it is not easy to distinguish with the naked eye on the electrophoresis bands, so it is not suitable for quickly and accurately identifying the purity of the seeds. InDel markers with stable amplification and large band differences are the screening targets.

Embodiment 2

[0032] Example 2: Using InDel molecular marker primers to identify the hybridization purity of Modilong wax gourd

[0033] (1) DNA extraction

[0034] 1. Weigh 8g (about 150 grains) of Modilong wax gourd seeds into a small nylon bag, soak the seeds in warm water at 55°C for 30 minutes, and stir continuously during the soaking process to ensure that the seeds are heated evenly and avoid parts that are not soaked. , wash off the mucus on the outer surface of the seeds with clean water, then place the seeds in clean water at room temperature for 8-12 hours, drain the water for later use;

[0035] 2. Disinfect the germination box with 0.1% mercuric chloride for 8-10 minutes, rinse it with clean water and set it aside. Lay two layers of germination test paper in the germination box and add water to keep it moist. Distribute the above seeds evenly in the germination box for germination. The temperature is set at 28-30°C, and distilled water is added to the germination box every day...

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Abstract

The invention discloses an InDel molecular marker primer of Modilong white gourds and an application of the InDel molecular marker primer. The primer comprises primer pairs corresponding to 7 point sites, and the nucleotide sequences are shown as SEQID NO.1-SEQ ID NO.14. When the Modilong white gourds are subjected to hybrid purity identification, firstly genome DNA of seeds of samples to be detected is extracted to be used as a template, then the primer is used for performing PCR amplification, and finally amplification products are detected through electrophoresis. Banding patterns of parents and hybrids are compared, the proportion of the number of real hybrids to the total number of seeds of strips which can be observed, namely the hybridization purity, is obtained. The InDel marker provided by the invention can realize effective distinguishing of the parents from the hybrids, the identification cost is low, the efficiency is high, and the coincidence degree of an identification result and a field identification result reaches 99% or above.

Description

technical field [0001] The invention relates to the technical field of molecular markers, in particular to a set of InDel molecular marker primers and applications thereof. Background technique [0002] Molecular markers are a new form of genetic markers developed after morphological markers, cell markers and biochemical markers. It is based on mutations in proteins and nucleic acid molecules, and detects biological genetic structures and their variations. There are many defects and problems in the identification of enzymes and proteins by electrophoresis, so they are widely used in genetic breeding, gene positioning and diversity analysis. Molecular marker methods commonly used in plant genetic research include restriction fragment length polymorphism (RFLP), random amplified polymorphism (RAPD) and simple repeat sequence marker (SSR), but the above-mentioned techniques have the following defects: RFLP requires The participation of radioactive isotopes is more complicated;...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156C12Q2600/13
Inventor 弭宝彬周火强谢玲玲武芳芳肖伟
Owner HUNAN VEGETABLE RES INST
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