Rapid molecule detection method of Pinus massoniana Lamb. and Pinus elliottii, and specific primer pair thereof

A technology for molecular detection and slash pine, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve problems affecting masson pine and slash pine, and achieve 100% accurate and guaranteed detection results The reliability and economic benefits are obvious

Inactive Publication Date: 2013-06-26
GUANGXI FORESTRY RES INST
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  • Claims
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AI Technical Summary

Problems solved by technology

[0004] Hybrid identification at the level of molecular genetics is one of the most reliable identification methods at present. However, there is no molecular biological identification method for masson pine, slash pine and their hybrids, which affects the smooth research on interspecific hybridization of masson pine and slash pine. conduct

Method used

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  • Rapid molecule detection method of Pinus massoniana Lamb. and Pinus elliottii, and specific primer pair thereof
  • Rapid molecule detection method of Pinus massoniana Lamb. and Pinus elliottii, and specific primer pair thereof
  • Rapid molecule detection method of Pinus massoniana Lamb. and Pinus elliottii, and specific primer pair thereof

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Experimental program
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Embodiment 1

[0027] Collection of materials

[0028] Take the needles of slash pine or masson pine, store them in ice packs (mainly to prevent the degradation of leaf tissue and browning of leaves), bring them back in foam plastic boxes, and store them in a -70°C refrigerator.

[0029] CTAB—silica bead method to extract DNA

[0030] Add 4ml of CTAB extract and 80ul of mercaptoethanol to a 10ml centrifuge tube (eppendorf tube), and preheat it in a 65°C water bath. Take 1-2g of the material in a mortar, add an appropriate amount of liquid nitrogen to quickly grind it several times into a fine powder, transfer it to the preheated CTAB extract, and seal the centrifuge tube with a sealing film to prevent the volatilization of mercaptoethanol. Put it back in the water bath and keep it warm for 30 minutes. During the keep warm process, shake it a few times to keep it even. Remove the Eppenedorf tube. Put it on ice and cool it down to room temperature quickly, add an equal volume of 4ml of chl...

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Abstract

The invention discloses a rapid molecule detection method of Pinus massoniana Lamb. and Pinus elliottii, and a specific primer pair thereof. The primer pair is PJ164L/R and has nucleotide sequences which are represented by SEQ. ID. No.1 and SEQ. ID. No.2 respectively in a sequence table, and the primer pair allows a 303bp product to be specifically amplified in Pinus massoniana Lamb. and a 325bp product to be specifically amplified in Pinus elliottii. The rapid molecule detection method of Pinus massoniana Lamb. and Pinus elliottii is established depended on the strong specificity characteristic of the sequences, allows determination to be carried out through the once implementation of a PCR amplification reaction and gel electrophoresis directly according to the fragment of the product, has the advantages of simple operation, accuracy, reliability and high sensitivity, is suitable for the rapid and reliable molecule detection and identification of Pinus massoniana Lamb. and Pinus elliottii, and has great scientific research values to identify the Pinus massoniana Lamb. and Pinus elliottii hybrid seedling in the future.

Description

technical field [0001] The invention relates to the identification of masson pine, slash pine and hybrid varieties thereof, in particular to a rapid molecular detection method of masson pine and slash pine and a pair of specific primers. Background technique [0002] Masson pine (Pinus massoniana Lamb.) is the most widely distributed coniferous tree species in my country. It is also an important tree species for timber, barren hills afforestation and industrial raw materials in southern China. Its wood and pine resin are the main raw materials for many forest industries, forestry industries and paper industries. Masson pine is distributed in the subtropical regions of my country (accounting for about 1 / 5 of the country's land) in the mountains. The total forest area ranks first among conifer species in the country, and its stock volume is second only to spruce, fir and larch, ranking fourth among conifer species in the country. Slash pine (Pinus elliottii) is one of the intr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 贾婕冯源恒杨章旗黄永利黄雪芬
Owner GUANGXI FORESTRY RES INST
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