Barnea dilatata mitochondrial COI gene amplification primer

A wide-shelled whole sea bamboo shoots, amplification primer technology, applied in genetic engineering, plant genetic improvement, botanical equipment and methods and other directions, can solve problems such as unsatisfactory amplification results, reduce the time and cost of amplification, Highly specific and efficient effects

Inactive Publication Date: 2016-04-06
MARINE BIOLOGY INST OF SHANDONG PROVINCE
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  • Application Information

AI Technical Summary

Problems solved by technology

Because the general primers COIL1490 and COIL2198 designed by Folmer (1994) had unsatisfactory amplification results in the w

Method used

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  • Barnea dilatata mitochondrial COI gene amplification primer
  • Barnea dilatata mitochondrial COI gene amplification primer
  • Barnea dilatata mitochondrial COI gene amplification primer

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0017] Example 1: Amplification and sequence analysis of the COI gene fragment of the whole sea bamboo shoot and several other shellfish genome COI gene fragments

[0018] Amplification of the COI gene of the wide-shelled whole sea bamboo shoot:

[0019] (1) DNA extraction was performed on wide-shelled whole sea bamboo shoots collected from Ningbo and Qinhuangdao, Zhejiang Province (16 in total, 32 in total) by CTAB method, and diluted with double distilled water to a working solution of 100 ng / μl for later use.

[0020] (2) Use the above primers to perform PCR amplification on the DNA working solution. The preparation solution of the reaction system includes: 1 μl template DNA, 0.05 μl Taq polymerase, 1 μl 10×PCRb μffer, 0.8 μl NTPs, 1 μl each of the upstream and downstream primers, 5.15 μl double-distilled water. The PCR amplification conditions were: pre-denaturation at 94°C for 3 min, denaturation at 94°C for 45 s, annealing temperature at 46-56°C for 45 s, extension at 7...

Embodiment 2

[0023] Embodiment 2: the design of primer

[0024] (1) DNA extraction was performed on wide-shelled whole sea bamboo shoots collected from Ningbo and Qinhuangdao, Zhejiang Province (16 in total, 32 in total) by CTAB method, and diluted with double distilled water to a working solution of 100 ng / μl for later use.

[0025] (2) Using COI universal primers (LCO1490F and HCO2198R, Folmer1994) to amplify the partial sequence of the whole sea bamboo shoot with broad shell.

[0026] (3) From the part of the wide-shell whole sea bamboo sequence obtained above, use the software BioEdit (http: / / www.Mbio.Ncsu.edu / BioEdit / bioedit.html) to perform multiple comparative analysis on the above sequence to determine the sequence before and after Conserved area.

[0027] (4) After comparison, 150 base pairs at the front and rear ends are in the conserved region, and 3 pairs of primers were designed with the primer design software PrimerPremier5. The primers designed were as far as possible witho...

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Abstract

The invention provides a barnea dilatata mitochondrial COI gene. The nucleotide sequence of the gene is SEQ ID NO: 1. The base sequence is used for designing an amplification primer for detecting barnea dilatata. The primer is designed for barnea dilatata mitochondrial COI gene segments. Compared with a COI gene primer universal for mollusks, the amplification primer has the advantage of absolute specificity. The amplification efficiency of the primer reaches as high as 90% or above, amplification efficiency is high, and time and cost for amplification are greatly reduced.

Description

technical field [0001] The invention belongs to the technical field of molecular marker screening, and in particular relates to a wide-shell whole sea bamboo shoot mitochondrial COI gene amplification primer. Background technique [0002] Wide-shelled whole sea bamboo shoots (Barneadilatata) belong to the mollusca (Mollusca), bivalve (Bivalvia), sea roaches (Myoida), sea bamboo shoots (Pholadidae), whole sea bamboo shoots (GenreBarnea Leach), distributed in the Philippines, Japan and my country's coastal areas; most of them live latently in the soft mud bottom near the river mouth, and can dive deep into the mud. Wide-shell whole sea bamboo shoots are shellfish distributed along the coast of China. They are rich in protein and vitamins, and the meat is delicious. There is artificial breeding in Fujian, Zhejiang and other places in my country. There have been many studies on the artificial propagation, reproductive growth, physiological and biochemical aspects of the whole s...

Claims

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Application Information

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IPC IPC(8): C12N15/53C12Q1/68C12N15/11
CPCC12N9/0053C12Q1/6888C12Y109/03001
Inventor 张婷婷李莉王英俊邹琰刘童刘洪军王慧
Owner MARINE BIOLOGY INST OF SHANDONG PROVINCE
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