PCR (polymerase chain reaction) primer group for species identification of cannabins satival, boehmeria nivea and linum usitatissimum fibers and PCR identification method

A primer set and ramie technology are applied in the PCR primer set for identifying hemp, ramie, and flax raw hemp fiber types, and in the field of PCR identification, which can solve problems such as difficulties and achieve the effects of reliable results, small sample size, and convenient detection.

Active Publication Date: 2016-01-27
保琦蓓
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the Japanese fiber identification standard, infrared spectroscopy is used to identify ramie and flax. In the Korean fiber identification standard, sisal is identified by the form of burning residue of hemp fiber. The Korean standard also uses fiber rotation direction and coloring reaction to identify hemp, flax, and jute. In actual operation, it is found that when using rotation direction and coloring reaction to identify hemp, flax and jute, it is quite difficult even for experienced inspectors without relevant reference objects
To further completely distinguish the hemp fibers of hemp, ramie, and flax through objective experimental detection, there is no literature report yet.

Method used

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  • PCR (polymerase chain reaction) primer group for species identification of cannabins satival, boehmeria nivea and linum usitatissimum fibers and PCR identification method
  • PCR (polymerase chain reaction) primer group for species identification of cannabins satival, boehmeria nivea and linum usitatissimum fibers and PCR identification method
  • PCR (polymerase chain reaction) primer group for species identification of cannabins satival, boehmeria nivea and linum usitatissimum fibers and PCR identification method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029]Embodiment 1: primer set described in the present invention

[0030] Hemp detection upstream and downstream primers: see SEQ ID NO:1 for the upstream primer, specifically 5′-ATCGAAGAGCAGCAAACAAGC-3′, see SEQ ID NO:2 for the downstream primer, specifically 5′-TGGAGATCTTGTCTTCCAGCTG-3′;

[0031] Ramie detection upstream and downstream primers: see SEQIDNO:3 for the upstream primer, specifically 5'-TCCAAGCATTTCCCGAACGA-3', see SEQIDNO:4 for the downstream primer, specifically 5'-GCCCGATCTTCCAGCTCTAC-3';

[0032] Flax detection upstream and downstream primers: see SEQ ID NO:5 for the upstream primer, specifically 5′-TCATCAACAGCGCATCTGGT-3′, and refer to SEQ ID NO:6 for the downstream primer, specifically 5′-AGCCTTACAGCCACACACTC-3′.

Embodiment 2

[0033] Embodiment 2: PCR identification method of the present invention

[0034] Adopt commercially available commercial DNA extraction kit to extract the DNA of the hemp plant fiber sample to be tested;

[0035] Adopt the nucleotide sequence shown in SEQIDNO:1-2 or the nucleotide sequence shown in SEQIDNO:3-4 or the nucleotide sequence shown in SEQIDNO:5-6 to carry out PCR amplification to sample DNA;

[0036] The PCR amplification system is:

[0037] The total reaction volume is 25μl, including 20ng sample DNA, 12.5μl 2×PremixTaq buffer, 1μl each of 5μM upstream and downstream primers, and the balance is ddH 2 O.

[0038] The procedure for PCR amplification is:

[0039] Pre-denaturation at 95°C for 5 minutes;

[0040] Denaturation at 95°C for 30s, annealing at 55°C for 30s, extension at 72°C for 45s, cycled 27 times;

[0041] Extend at 72°C for 10min, and store at 10°C.

[0042] PCR amplified product carries out agarose gel electrophoresis (gel concentration 2.0%), if ...

Embodiment 3

[0043] Embodiment 3: identification test

[0044] The test samples were hemp, ramie and flax, and the specific varieties and collection places are shown in Table 1.

[0045] Table 1 Test sample

[0046] sample number

breed name

collection place

A (hemp)

Yunma No.1

Xishuangbanna, Yunnan Province

B (hemp)

Heilongjiang industrial hemp

Mohe, Heilongjiang Province

C (hemp)

Anhui Cannabis

Lu'an, Anhui Province

D (ramie)

Xiangzhu No.2

Changsha, Hunan

E (ramie)

Xiangzhu No.3

Changsha, Hunan

F (flax)

West Asia One

Guyuan, Ningxia Province

G (linen)

Gia II

Changchun City, Jilin Province

H (linen)

Gia IV

Changchun City, Jilin Province

I (linen)

Gia Six

Changchun City, Jilin Province

[0047] Adopt the nucleotide sequence shown in SEQIDNO:1-2 to carry out PCR amplification to above-mentioned sample DNA respectively and carry ou...

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Abstract

The invention relates to the technical field of biological information and discloses a PCR (polymerase chain reaction) primer group for species identification of cannabins satival, boehmeria nivea and linum usitatissimum fibers and a PCR identification method. The primer group is formed by nucleotide sequences shown as SEQ ID NO:1-6, wherein the nucleotide sequences shown as SEQ ID NO:1-2 are sense and antisense primers for identifying cannabins satival, the nucleotide sequences shown as SEQ ID NO:3-4 are sense and antisense primers for identifying boehmeria nivea, and the nucleotide sequences shown as SEQ ID NO:5-6 are sense and antisense primers for identifying linum usitatissimum. The primer group high in specificity is provided by means of biotechnology and can be used for exclusive identification of the cannabins satival, the boehmeria nivea and the linum usitatissimum without mutual identification interference. In addition, by the aid of the PCR technology, the method has the advantages of small sample size, convenience and stability in detection, high operability, result reliability and the like and is widely applicable to detection and component analysis of bast fibers for textiles.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a PCR primer set and a PCR identification method for identifying the types of hemp, ramie and flax raw hemp fibers. Background technique [0002] Hemp (Cannabinssatival L.) is an annual herb of the genus Cannabis of the family Cannabis, also known as industrial hemp, hemp, and is an important bast fiber crop. At present, the cultivated area and output are mostly in China, mainly distributed in Yunnan, Anhui, Heilongjiang and other places in China. At present, my country is still the main producer of hemp fiber, and its output accounts for about 1 / 3 of the world's hemp output, ranking first in the world. Hemp fiber has become the green new textile and clothing material with the most sustainable development potential in the 21st century due to its excellent environmental protection and unique antibacterial comfort. Ramie (Boehmerianivea L. Gand) is a perennial herbaceous plant of the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 保琦蓓傅科杰任清庆冯云尤建武杨力生
Owner 保琦蓓
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