Method and PCR (polymerase chain reaction) primer for identifying dendrobium officinale

A technology of Dendrobium officinale and Dendrobium candidum, applied in the field of PCR primers, can solve problems such as poor stability, difficulty in grasping and teaching, and achieve the effects of low requirements, avoiding false identification, and sensitive identification methods

Inactive Publication Date: 2017-03-22
三明市农业科学研究院
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Problems solved by technology

Traditional identification based on traits and microscopic features is difficult to grasp and teach in practice due to the influence of the experience of the identifyrs, indi

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  • Method and PCR (polymerase chain reaction) primer for identifying dendrobium officinale
  • Method and PCR (polymerase chain reaction) primer for identifying dendrobium officinale

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[0034] The specific embodiments of the present invention will be described below with reference to the drawings.

[0035] In this specific embodiment, the samples to be tested are wild Dendrobium officinale collected from Taining, Fujian Province, Sanming Academy of Agricultural Sciences, and wild Dendrobium candidum collected from Huyuan Township, Shaxian County, Fujian Province. The above materials are identified by Professor Wei Daozhi of Fujian Agriculture and Forestry University. The original plant of Dendrobium officinale Kimura et Migo and Dendrobium moniliforme (L.) Sw.).

[0036] A method for identifying Dendrobium officinale includes the following steps:

[0037] 1. Extract the DNA of the sample to be tested;

[0038] The modified CTAB method was used to extract the total DNA from the leaves of Dendrobium candidum and Dendrobium tenuiflorum.

[0039] Take 0.1g of young leaves, add liquid nitrogen to grind to powder, transfer to a 2mL centrifuge tube; add 800μL of 70℃ preheate...

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Abstract

The invention relates to a method and a PCR (polymerase chain reaction) primer for identifying dendrobium officinale. The method includes identifying the dendrobium officinale on the basis of mononucleotide difference sites of chloroplast ACCD gene sequences of the dendrobium officinale and other herba dendrobii of dendrobium by the aid of Tetra-primer ARMS-PCR processes. The PCR primer for identifying the dendrobium officinale comprises specific inner primers for identifying the dendrobium officinale and the other herba dendrobii of the dendrobium and a pair of specific outer primers. The method is simple, and the dendrobium officinale can be detected by the aid of one-step PCR and conventional electrophoretic techniques; identification results are accurate, mismatched basic groups are introduced when the primers are designed, accordingly, the specificity of PCR amplification reaction can be greatly enhanced, and false negative results can be prevented owing to positive control of outer primer strips; further, the method is sensitive, and the dendrobium officinale can be completely identified only by a few samples.

Description

technical field [0001] The invention relates to a method for identifying dendrobium officinale based on a single nucleotide difference site in a chloroplast DNA sequence, in particular to a method for identifying dendrobium officinale and a PCR primer used therefor. Background technique [0002] Dendrobium officinale (Dendrobium officinale), also known as black knot grass, is a perennial epiphytic herb of Orchidaceae (Orchidaceae). They are mainly distributed in Guangxi, Yunnan, Zhejiang, Fujian, Anhui and other places in my country. Dendrobium candidum is known as "life-saving fairy grass" and "inch golden grass" in the folks. It has remarkable therapeutic and health effects. Because Dendrobium candidum is very rare, due to human over-excavation and serious habitat damage, coupled with the unique habitat of Dendrobium candidum and difficult reproduction, As early as 1987, Dendrobium officinale was included in the "List of National Key Protected Wild Medicinal Species". Due ...

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2600/156C12Q2545/101
Inventor 曹奕鸯叶炜江金兰李永清范志敏雷伏贵杨学周建金莫智龙颜沛沛
Owner 三明市农业科学研究院
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