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Simple sequence repeat (SSR) marker polymerase chain reaction (PCR) reaction method applied to chrysanthemum and related species thereof universally

A technology of labeling, chrysanthemum, applied in the field of plant biology, to achieve the effect of easy operation, good repeatability and good versatility

Active Publication Date: 2013-01-16
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, in Chrysanthemum and its close relatives (Chrysanthemum, Chrysanthemum, Taihang Chrysanthemum, and Hibiscus Chrysanthemum), an efficient and versatile PCR reaction method for SSR markers has not been developed.

Method used

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  • Simple sequence repeat (SSR) marker polymerase chain reaction (PCR) reaction method applied to chrysanthemum and related species thereof universally
  • Simple sequence repeat (SSR) marker polymerase chain reaction (PCR) reaction method applied to chrysanthemum and related species thereof universally
  • Simple sequence repeat (SSR) marker polymerase chain reaction (PCR) reaction method applied to chrysanthemum and related species thereof universally

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Effect test

Embodiment 1

[0026] Embodiment 1 The establishment of the common SSR marker PCR reaction method of Chrysanthemum and its related species in the present invention

[0027] (1) DNA samples of commonly used wild species and their cultivated varieties in Chrysanthemum, Chrysanthemum, Taihang Chrysanthemum, and Hibiscus were extracted.

[0028] The wild species of chrysanthemum are: wild chrysanthemum, chamomile, chamomile, Maohua chrysanthemum, small red chrysanthemum, chrysanthemum brain, purple wild chrysanthemum, and Shennong chrysanthemum.

[0029] Chrysanthemum cultivars include: Sunshine Chrysanthemum (potted chrysanthemum), Fanbailu (northern ground cover chrysanthemum), Sijihuang (northern ground cover chrysanthemum), Xiangyu (cut flower chrysanthemum), Shanghai chrysanthemum (southern ground cover chrysanthemum), Xiaju (summer flowering variety), Guoqinghong (small chrysanthemum in southern ground cover), Yurenmian (chrysanthemum variety for tea);

[0030] The chrysanthemums in Chrys...

Embodiment 2

[0063] Embodiment 2 utilizes the method of the present invention to carry out the screening of chrysanthemum SSR marker

[0064] (1) According to the design principles of SSR molecular markers, according to the published EST sequence of chrysanthemum, use the Primer5 design software to design 3 pairs of SSR primers:

[0065] SSR marker YH-1 primer sequence:

[0066] Upstream primer: 5'-TTCGCATGGGTGGTGGGTT-3' (as shown in SEQ ID NO.5)

[0067] Downstream primer: 5'-CAACAACACAACATGGGGG-3' (as shown in SEQ ID NO.6)

[0068] SSR marker YH-2 primer sequence:

[0069] Upstream primer: 5'-TTCGACGGGTGGTGGTGTT-3' (as shown in SEQ ID NO.3)

[0070] Downstream primer: 5'-GACACCACAACGCTACCGC-3' (as shown in SEQ ID NO.4)

[0071] SSR marker YH-3 primer sequence:

[0072] Upstream primer: 5'-GGGACCCAAGCAGTAGAATG-3' (as shown in SEQ ID NO.7)

[0073] Downstream primer: 5'-ACCAAACAACCCTCCAACC-3' (as shown in SEQ ID NO.8)

[0074] (2) Extract DNA samples of chrysanthemum varieties Yulon...

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Abstract

The invention discloses a simple sequence repeat (SSR) marker polymerase chain reaction (PCR) reaction method universally applied to chrysanthemum and related species thereof. In a PCR reaction system, the primer concentration is 0.32 MuM / Mul; the Taq enzyme concentration is 0.2 U / Mul; the dNTP concentration is 0.4 MuM / Mul; the Mg<2+> concentration is 0.65 MuM / Mul; and the DNA concentration is 2 to 4 ng / Mul. Stripes obtained by the method for performing SSR molecular marker PCR reaction are clear and have high polymorphism and high repeatability. The method is simple in operation and commonly used in wild species and varieties of chrysanthemum, Ajania, Opisthopappus Shih and Crossostephium Less, and has a wide application prospect.

Description

technical field [0001] The invention belongs to the field of plant biology technology, and in particular relates to a common SSR marker PCR reaction method for Chrysanthemum and its related species. Background technique [0002] Polymerase Chain Reaction (PCR) is an in vitro nucleic acid amplification technique developed in the mid-1980s. It has the advantages of specificity, sensitivity, high yield, rapidity, simplicity, good repeatability, and easy automation. It can amplify the target gene or a certain DNA fragment to be studied to hundreds of thousands or even hundreds of thousands within a few hours in a test tube. Tens of thousands of times, so that the naked eye can directly observe and judge. [0003] Five factors of PCR: primers, TaqDNA polymerase, dNTP, DNA template and Mg 2+ . [0004] The basic principle of PCR technology: similar to the natural replication process of DNA, its specificity depends on oligonucleotide primers complementary to both ends of the tar...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 张启翔刘华孙明程堂仁王佳潘会堂
Owner BEIJING FORESTRY UNIVERSITY
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