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Molecular-specific marker primers and detection methods for hibiscus varieties Brick, Norwood No. 1, Sanchoyo and Norwood No. 2

A technology for labeling primers and detection methods, which is applied in biochemical equipment and methods, microbe determination/testing, DNA/RNA fragments, etc. It can solve the problems that DNA molecular marker technology has not yet been widely used in genetic diversity, and achieve simple methods Effect

Active Publication Date: 2019-09-20
ZHEJIANG FORESTRY ACAD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the DNA molecular marker technology that has been widely used in recent years has not been widely used in the classification of hibiscus varieties, the relationship between varieties and genetic diversity.

Method used

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  • Molecular-specific marker primers and detection methods for hibiscus varieties Brick, Norwood No. 1, Sanchoyo and Norwood No. 2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] (1) Extraction of genomic DNA of hibiscus varieties:

[0043] Take 0.01 g of the young leaves of the hibiscus species to be tested, add liquid nitrogen and grind them thoroughly, and use the SDS-CTAB method to extract the genomic DNA. The crude DNA was purified by Magabio Nucleic Acid Purification Kit (Bioer, Hangzhou, China), and detected by 1.5% agarose gel electrophoresis and DNA / RNA UV spectrophotometer (GeneQuant Pro, GE Healthcare). Integrity, Purity and Concentration. DNA samples with OD260 / OD280>1.8 were used for subsequent PCR amplification. DNA extracts were stored in a -20°C refrigerator for later use.

[0044] (2) Design specific PCR amplification primers, the sequence of the primer pair is:

[0045] Upstream primer: 5′-CTCTCTCAACTCCCTCATCGG-3′;

[0046] Downstream primer: 5′-GGAAAAAAAACAAAAAACAAAAG-3′;

[0047] Sangon Bioengineering (Shanghai) Co., Ltd.

[0048] (3) PCR amplification of SCAR molecular markers:

[0049] Composition of PCR reaction sol...

Embodiment 2

[0055] (1) Extraction of genomic DNA of hibiscus varieties:

[0056] Take 0.02 g of young leaves of the hibiscus species to be tested, add liquid nitrogen and grind them thoroughly, and use the SDS-CTAB method to extract the genomic DNA. The crude DNA was purified by Magabio Nucleic Acid Purification Kit (Bioer, Hangzhou, China), and detected by 1.8% agarose gel electrophoresis and DNA / RNA UV spectrophotometer (GeneQuant Pro, GE Healthcare). Integrity, Purity and Concentration. DNA samples with OD260 / OD280>1.8 were used for subsequent PCR amplification. DNA extracts were stored in a -22°C refrigerator for later use.

[0057] (2) Design specific PCR amplification primers, the sequence of the primer pair is:

[0058] Upstream primer: 5′-CTCTCTCAACTCCCTCATCGG-3′;

[0059] Downstream primer: 5′-GGAAAAAAAACAAAAAACAAAAG-3′;

[0060] Sangon Bioengineering (Shanghai) Co., Ltd.

[0061] (3) PCR amplification of SCAR molecular markers:

[0062] Composition of PCR reaction solutio...

Embodiment 3

[0068] (1) Extraction of genomic DNA of hibiscus varieties:

[0069] Take 0.01 g of the young leaves of the hibiscus species to be tested, add liquid nitrogen and grind them thoroughly, and use the SDS-CTAB method to extract the genomic DNA. The crude DNA was purified by Magabio Nucleic Acid Purification Kit (Bioer, Hangzhou, China), and detected by 1.6% agarose gel electrophoresis and DNA / RNA UV spectrophotometer (GeneQuant Pro, GE Healthcare). Integrity, Purity and Concentration. DNA samples with OD260 / OD280>1.8 were used for subsequent PCR amplification. DNA extracts were stored in a -18°C refrigerator for later use.

[0070] (2) Design specific PCR amplification primers, the sequence of the primer pair is:

[0071] Upstream primer: 5′-CTCTCTCAACTCCCTCATCGG-3′;

[0072] Downstream primer: 5′-GGAAAAAAAACAAAAAACAAAAG-3′;

[0073] Sangon Bioengineering (Shanghai) Co., Ltd.

[0074] (3) PCR amplification of SCAR molecular markers:

[0075]Composition of PCR reaction solu...

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Abstract

The invention relates to a molecular specific marker primer and a detection method for Hibiscus varieties of Hibiscus syriacus 'Bricutts', Hibiscus syriacus 'Notwoodone', Hibiscus syriacus 'Sanchoyo' and Hibiscus syriacus 'Notwoodtwo', which are applied to identification of the Hibiscus varieties. The Hibiscus varieties are widely used in gardens, but are difficult to identify from the aspectof phenotypic characteristics. The molecular specific marker primer sequences of the Hibiscus varieties of Hibiscus syriacus 'Bricutts', Hibiscus syriacus 'Notwoodone', Hibiscus syriacus 'Sanchoyo'and Hibiscus syriacus 'Notwoodtwo' are: upstream primer: 5'-CTCTCTCAACTCTCATCGG-3'; downstream primer: 5'-GGAAAAAAAAAAAACA-AAAG-3'. The molecular specific marker primer can be applied to rapid earlyidentification of the Hibiscus varieties of Hibiscus syriacus 'Bricutts', Hibiscus syriacus 'Notwoodone', Hibiscus syriacus 'Sanchoyo' and Hibiscus syriacus 'Notwoodtwo'. The method is simple, fastand accurate, and is an irreplaceable molecular means for identifying Hibiscus varieties by apparent characteristics.

Description

technical field [0001] The invention relates to a molecular specific marker primer and a detection method of hibiscus varieties Brick, Norwood No. 1, Sanchoyo and Norwood No. 2, which are used for identification of hibiscus varieties. Background technique [0002] Hibiscus belongs to the genus Hibiscus of Malvaceae and is a world-renowned ornamental plant in gardens. At present, there are more than 200 varieties of hibiscus known. Because of its beautiful plant shape, bright colors and long flowering period, it has great application value in gardens. At present, on a global scale, the research and cultivation of hibiscus species are mainly focused on ornamental resources. In recent years, attention has been paid to research as a potential new resource for edible flowers and natural food colorants, among other uses. In terms of breeding and cultivation of new varieties of edible flowers, there are farms and households specialized in growing edible flowers abroad, such as th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
Inventor 杨少宗方茹刘本同徐梁林昌礼程亚平
Owner ZHEJIANG FORESTRY ACAD
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