SSR multiple detection primer, kit and method for pithecellobium clypearia variety identification

A variety identification and multiple detection technology, which is applied in biochemical equipment and methods, microbial measurement/inspection, recombinant DNA technology, etc., can solve the problem that there is no multiple SSR detection technology system for monkey earring variety identification, and save experimental costs , Reagent saving, good economical effect

Pending Publication Date: 2022-04-05
RES INST OF TROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Unfortunately, there is no efficient multiple SSR detection technology system for the identification of monkey earring varieties, which needs to be developed and used for effective identification of monkey earring varieties

Method used

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  • SSR multiple detection primer, kit and method for pithecellobium clypearia variety identification
  • SSR multiple detection primer, kit and method for pithecellobium clypearia variety identification
  • SSR multiple detection primer, kit and method for pithecellobium clypearia variety identification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] (1) The 15 SSR markers of monkey earrings are optimized into 7 combined triple or double detection technology systems

[0045] Based on the 456 SSR-labeled primer sequences developed by our laboratory in the early stage, taking into account that the primers do not interfere with each other (not paired with each other), the PCR amplification program is the same, the length range of the amplified fragment is different and the concentration of the amplified product (the peak height of the detection ) and other factors, select 15 highly polymorphic markers with polymorphic information content (polymorphic information content, PIC) higher than 0.5 and optimize them as 7 combinations of triple or double detection, and the 5' end of the forward primer of the same combination Modified with different fluorophores (FAM, HEX, ROX or TAM); the specific 7 combinations (the asterisks indicate primer-modified fluorophores) are:

[0046] Marker combination A: ARCeSSR266(*HEX), ARCeSSR1...

Embodiment 2

[0078] Example 2: Identification of 20 varieties (cutting clones) of monkey earrings to be tested based on SSR molecular markers

[0079] (1) DNA extraction of monkey earring species. The 20 varieties to be inspected still come from Huadu Cutting Nursery of Guangdong Province (23°24'12"N, 113°13'06"E). About 3g of young leaves of each variety are collected, which can be temporarily refrigerated or long-term frozen.

[0080] DNA extraction from leaf samples was performed using a modified CTAB (cetyltrimethylammonium bromide) method. Concrete method is with (2) step (1) among the embodiment 1.

[0081] (2) Multiplex polymerase chain reaction (PCR) and detection of 15 SSR molecular markers. The 15 SSR molecular markers optimized based on the present invention are divided into 7 groups of triple or double detection techniques (marker grouping and primer modification fluorescence are the same as (1) in Example 1), using the DNA extracted in step (1) as a template, Perform PCR amp...

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Abstract

The invention discloses SSR multiple detection primers, a kit and a method for pithecellobium clypearia variety identification. Compared with independent PCR detection of a single marker, the method has the advantages of good economical efficiency, reagent saving, time saving, high efficiency and the like, and a simple and reliable method is provided for population diversity analysis of pithecellobium clypearia and sibling species, genetic evaluation of germplasm resources, authenticity identification of varieties and medicinal materials and the like; the method is used for construction of SSR molecular fingerprints of pithecellobium clypearia varieties (cutting clones) and identification of to-be-detected varieties, has the advantages of being accurate, reliable, effective and rapid, can effectively discriminate counterfeit and disordered varieties and new varieties, and provides important technical support for guaranteeing rights and interests of improved variety cultivators and forest planters.

Description

technical field [0001] The invention belongs to the technical field of molecular markers, and in particular relates to SSR multiple detection primers, a kit and a method for monkey earring variety identification. Background technique [0002] Monkey earring [Archidendron clypearia (Jack) I.C.Nielsen] is an important medicinal tree species of Leguminosae Juss. (Caesalpinioideae DC.) monkey earring (Archidendron F.Mueller), naturally distributed in tropical Asia and subtropical Asia area. The industrial utilization value of monkey earrings is extremely high. The extracts of its branches and leaves can be used to treat diseases such as upper respiratory tract infection, acute pharyngitis and tonsillitis. The dregs of the medicine can be used to prepare high-density composite materials. The natural regeneration ability of monkey earrings is weak, coupled with the serious interference of human activities, wild resources are becoming increasingly scarce, and the existing natural ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/686C12N15/11
Inventor 李梅李丹丹翁启杰周明明周长品甘四明
Owner RES INST OF TROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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