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Primers, detection method and application for diploid Haynaldia villosa 3V chromosome specific KASP marker detection

A technology of marker detection and detection method, which is applied in the field of molecular biology, can solve problems such as time-consuming and labor-intensive, and achieve the effects of improving efficiency, accelerating transfer and utilization, and accelerating the breeding process

Active Publication Date: 2021-02-26
CHENGDU INST OF BIOLOGY CHINESE ACAD OF S +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these markers are designed based on the polymorphism of the target fragment, and are mainly detected by PCR and electrophoresis, which is time-consuming and labor-intensive

Method used

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  • Primers, detection method and application for diploid Haynaldia villosa 3V chromosome specific KASP marker detection
  • Primers, detection method and application for diploid Haynaldia villosa 3V chromosome specific KASP marker detection
  • Primers, detection method and application for diploid Haynaldia villosa 3V chromosome specific KASP marker detection

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Embodiment 1

[0041] A preferred embodiment of the present invention provides a diploid 3V chromosome-specific KASP marker detection primer, the nucleotide sequence of the primer is as follows:

[0042] Upstream primer F1: 5'-GAAGGTGACCAAGTTCATGCTGAGCAGGCTGTCGAAGCTA-3';

[0043] Upstream primer F2: 5'-GAAGGTCGGAGTCAACGGATTGAGCAGGCTGTCGAAGCTC-3';

[0044] Downstream primer R: 5'-TGGAGCACGAGGGTGTGA-3'.

Embodiment 2

[0046] The present invention is based on the method for using the above-mentioned primers to detect the 3V chromosome of diploid T. villosa provided by the above-mentioned embodiment 1, comprising the following steps:

[0047] 1. Using the CTAB method to extract the genomic DNA of the plant to be tested, the extraction steps are as follows:

[0048] 1) Take 2g of fresh young leaves, grind them into fine powder with liquid nitrogen, add 2×CTAB extract (2% CTAB; 1.4M NaCl, 0.1M Tris-HCl, pH 8.0, 0.1M EDTA, pH 8.0) 15ml, mix well.

[0049] 2) 30-45min in a water bath at 65°C, during which time shake gently to mix. After cooling to room temperature, add an equal volume of chloroform:isoamyl alcohol (24:1), mix gently until the supernatant becomes milky, and centrifuge at 4000rpm for 10min.

[0050]3) Take the supernatant, add an equal volume of isopropanol, and place in an ice bath to precipitate DNA.

[0051] 4) The DNA was hooked out, washed twice with 70% ethanol and once wi...

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Abstract

The invention discloses primers, a detection method and application for diploid Haynaldia villosa 3V chromosome specific KASP marker detection. Nucleotide sequences of the primers for Haynaldia villosa 3V chromosome specific KASP marker detection are: F1: 5'-GAAGGTGACAAG TTCATGAGCTGAGCTGTCGAGCTAAGCTA-3'; F2: 5'-GAAGGTCGAGTCAGGACGATTGTCGAGGAGCTGTCAGCTGAGCTGAGCTCTCCTC-3'; and R: 5'-TGGAGCAGCGAGGTGTGTGA-3'. By utilizing the marker disclosed by the invention, the Haynaldia villosa 3V chromosome can be efficiently detected in a common wheat CS background, and a foundation is provided for applying excellent traits on the 3V chromosome in wheat breeding.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a primer, a detection method and an application for detecting a diploid 3V chromosome-specific KASP marker. Background technique [0002] SNP (single nucleotide polymorphism, single nucleotide polymorphism) refers to the DNA sequence polymorphism caused by the variation of a single base in the genomic DNA sequence. This variation is mainly caused by the conversion and transversion of a single base , extremely large in number, rich in polymorphism, dimorphic, easy to detect and count, and can realize high-throughput automated operation detection. With the maturity of next generation sequencing (NGS, next generation sequencing) technology, the development and detection of SNP has become easier and easier. Therefore, SNP molecular markers have been rapidly applied to various biological research fields and are regarded as the most important and most effective. ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/156C12Q2600/13C12Q2531/113C12Q2563/107Y02A50/30
Inventor 邓光兵龙海张洁蒋云王颖郭元林
Owner CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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