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Primer for detecting NCED gene of bupleurum chinense and real-time fluorescent quantitative PCR (polymerase chain reaction) method

A real-time fluorescent quantitative and fluorescent quantitative technology, applied in the field of fluorescent quantitative PCR

Pending Publication Date: 2022-04-15
HENAN UNIV OF CHINESE MEDICINE
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Problems solved by technology

However, there is no systematic report on the real-time fluorescent quantitative PCR detection method of the NCED gene in Bupleurum chinensis.

Method used

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  • Primer for detecting NCED gene of bupleurum chinense and real-time fluorescent quantitative PCR (polymerase chain reaction) method

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Embodiment 1

[0034] Such as figure 1 As shown, the present invention discloses a primer and a real-time fluorescence quantitative PCR method for detecting the NCED gene of Bupleurum chinensis, and the technical scheme adopted comprises the following steps:

[0035] Step 1, design of real-time fluorescent quantitative primers for the target gene NCED of Bupleurum chinensis:

[0036] Referring to the sequence (SEQ ID NO.1) of the NCED gene of Bupleurum chinensis, primers with suitable product fragment size and excellent parameters were designed. The sequence of the NCED gene of Bupleurum chinensis is shown in SEQ ID NO.1, which contains 1794 nucleotide bases Base, the nucleotide sequence shown in SEQ ID NO.1 is from the North Bupleurum NCED gene sequence obtained by the inventor's previous research; using Primer Premier5 software, referring to the North Bupleurum NCED gene sequence (SEQ ID NO.1), the product is designed Primer sequences with a fragment size of 80-200 bp and excellent parame...

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Abstract

The invention discloses a primer for detecting NCED genes of bupleurum chinense and a real-time fluorescent quantitative PCR (Polymerase Chain Reaction) method, belongs to the field of fluorescent quantitative PCR methods, and aims to solve the problem that a primer for detecting NCED genes of bupleurum chinense and a real-time fluorescent quantitative PCR method are lacked in the prior art. The NCED gene of the bupleurum chinense cannot be quickly, efficiently and highly sensitively detected. Total RNA of different tissues (roots, stems, leaves and flowers) of bupleurum chinense is extracted, an actin gene of bupleurum chinense is used as a reference gene for contrast, real-time fluorescent quantitative PCR detection is carried out, and distribution and expression differences of an NCED gene in different tissues (roots, stems, leaves and flowers) of bupleurum chinense are detected and analyzed by adopting a 2-delta delta Ct method. The real-time fluorescent quantitative PCR detection of the NCED genes of different tissues of bupleurum chinense can be realized, and the kit has the characteristics of rapidness, high efficiency and high sensitivity, and has important significance for researching the drought-resistant adversity stress related mechanism of bupleurum chinense.

Description

technical field [0001] The invention relates to the field of fluorescent quantitative PCR methods, in particular to a primer for detecting the NCED gene of Bupleurum chinensis and a real-time fluorescent quantitative PCR method. Background technique [0002] Bupleurum chinense DC. is a plant of the genus Bupleurum in the Umbelliferae family. Bupleurum chinense DC., one of the bases of Bupleurum chinense, is a bulk of Chinese medicinal materials in my country. At present, Bupleurum bupleurum is mainly distributed in arid and semi-arid areas in my country, with a large geographical span and a wide distribution range, and its growth and development are affected by the external environment. At each stage of growth and development, plants will regulate their own metabolism through different plant hormones, so as to adapt to changes in the external environment. Plant hormones such as abscisic acid participate in the regulation of the entire life cycle of plants, and play a very i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q2531/113C12Q2521/107C12Q2563/107C12Q2545/101
Inventor 杨林林初雷霞乔璐苏秀红董诚明
Owner HENAN UNIV OF CHINESE MEDICINE
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