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A kind of DNA molecular marker method of peony genome

A DNA molecule and labeling method technology, applied in recombinant DNA technology, DNA/RNA fragments, biochemical equipment and methods, etc., can solve problems such as low efficiency, achieve broad application prospects, reduce application difficulty, and simple application prospects.

Inactive Publication Date: 2016-01-13
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If you want to carry out this type of marking, you must first obtain the LTR sequence of the retrotransposon, and the efficiency of the current method for isolating the LTR sequence is still low

Method used

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  • A kind of DNA molecular marker method of peony genome
  • A kind of DNA molecular marker method of peony genome
  • A kind of DNA molecular marker method of peony genome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] A kind of peony genomic DNA molecular marker method is characterized in that: its specific operation steps are:

[0051] Step 1. Extraction of Peony Genomic DNA: Peony Genomic DNA was extracted using the improved CTAB method, and a total of 40 peony varieties were extracted (including Gejinzi, Wanhuasheng, Daomin, Luhefen, Huhong, Baiyu, Three Variations of Saiyu, Zhongshenghong, Green Fragrant Ball, Quhaohao, Erqiao, Oolong Pengsheng, Rouge Figure, Red Crown and Jade Belt, Meat Lotus, Silver and Red Pair, Peach Red, Jindao, Blue Sea and Bibo, Yu Jiyan Clothing, Shouxing Red, Xiaohu Hong, Silver Scale Jade Pearl, Red Lantern, Shengdan Furnace, Yao Huang, Pingshiyan, Arhat Red, Brocade Red, Red Lotus, Buddhist Cassock, Bingshan Jadeite, First Case Red, Number One Scholar Red , Rose Purple, Daojin, Dahuahuang, Purple Peony, Sichuan Peony, Yangshan Peony) genomic DNA;

[0052] Step 2. Digestion of Peony Genomic DNA: Use the MseI and EcoRI restriction endonucleases in the ...

Embodiment 2

[0058]A kind of peony genomic DNA molecular marker method is characterized in that: its specific operation steps are:

[0059] Step 1. Extraction of Peony Genomic DNA: Peony Genomic DNA was extracted using the improved CTAB method, and a total of 40 peony varieties were extracted (including Gejinzi, Wanhuasheng, Daomin, Luhefen, Huhong, Baiyu, Three Variations of Saiyu, Zhongshenghong, Green Fragrant Ball, Quhaohao, Erqiao, Oolong Pengsheng, Rouge Figure, Red Crown and Jade Belt, Meat Lotus, Silver and Red Pair, Peach Red, Jindao, Blue Sea and Bibo, Yu Jiyan Clothing, Shouxing Red, Xiaohu Hong, Silver Scale Jade Pearl, Red Lantern, Shengdan Furnace, Yao Huang, Pingshiyan, Arhat Red, Brocade Red, Red Lotus, Buddhist Cassock, Bingshan Jadeite, First Case Red, Number One Scholar Red , Rose Purple, Daojin, Dahuahuang, Purple Peony, Sichuan Peony, Yangshan Peony) genomic DNA;

[0060] Step 2. Digestion of Peony Genomic DNA: Use the MseI and EcoRI restriction endonucleases in the e...

Embodiment 3

[0066] A kind of peony genomic DNA molecular marker method is characterized in that: its specific operation steps are:

[0067] Step 1. The genomic DNA of peony was obtained by using the improved CTAB method to extract a total of 40 peony varieties (including Gejinzi, Wanhuasheng, Daomin, Luhefen, Huhong, Baiyu, Sanbian Saiyu, Zhong Raw Red, Green Fragrance Ball, Que Hao, Er Qiao, Oolong Holding Sheng, Rouge Figure, Red Crown and Jade Belt, Meat Hibiscus, Silver Red Delicate Pair, Peach Red Flattery, Golden Island, Blue Sea and Bibo, Concubine Concubine Dress, Longevity Red, Small Hu Hong, Silver Scale and Jade Pearl, Red Lantern, Sheng Danlu, Yao Huang, Ping Shiyan, Arhat Red, Brocade Pao Red, Red Lotus, Buddhist Cassock, Bingshan Emerald, First Case Red, Number One Scholar Red, Rose Purple, Island Brocade, Genomic DNA of Dahuahuang, Purple Peony, Sichuan Peony, Yangshan Peony);

[0068] Step 2. Digestion of Peony Genomic DNA: Use the MseI and EcoRI restriction endonucleases...

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Abstract

A method for molecular labeling of peony genome DNA, which comprises extracting peony genome DNA for enzyme digestion, linking the enzyme cleavage product with adapter DNA; performing PCR amplification on the ligation product and pre-amplification primers to obtain a pre-amplification product; taking the resulting pre-amplification The amplification product is amplified by PCR with the endonuclease linker selective amplification primer iPBS primer to obtain the PCR selective amplification product; then the PCR selective amplification product is taken, added to the staining solution and spotted for electrophoresis. After the electrophoresis is completed, Silver staining, observation, photography, and analysis of the results. The present invention is a molecular marker technology based on PCR and enzyme digestion, with simple operation, low cost and strong applicability; it is designed for peony genome research technology and develops a new retrotransposon-based molecular marker system, providing new technical means for the study of genetic variation and evolution of peony germplasm resources.

Description

technical field [0001] The invention relates to a molecular marker method, in particular to a molecular marker method of peony genome DNA. Background technique [0002] peony( Paeonia Suffruticosa Andr.) is Paeoniaceae (Paeoniaceae) genus Paeoniae ( Paeonia L.) Peony group (Sect. Mouton DC.) Deciduous shrub is an important ornamental greening plant and resource plant. Its flowers are beautiful in shape, bright in color and fragrant, and are praised by people of all ages. history. In the long-term cultivation and domestication process of Chinese peony, more than 1,000 cultivars with different petal colors, numbers, flower shapes and leaf shapes have been formed, represented by the Central Plains, Northwest, Jiangnan and Southwest varieties groups, and the cultivation areas are all over the country. everywhere. There are 9 species in the peony group (Hong Deyuan and Pan Kaiyu, 1999; Hong and Pan, 2007), distributed in 9 provinces including Henan, Gansu, Shaanxi, Shanxi,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6855C12Q1/686C12Q2525/191C12Q2531/113
Inventor 郭大龙侯小改段亚宾魏冬峰吕静霞
Owner HENAN UNIV OF SCI & TECH
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