Muricidae mitochondria 16s rRNA gene amplification primer

An amplification primer, mitochondrial technology, applied in DNA preparation, recombinant DNA technology and other directions, can solve the problems of large inter-species variability, low amplification efficiency of 16SrRNA non-specific amplification primers, and hindering the use of molecular technology.

Inactive Publication Date: 2013-04-24
OCEAN UNIV OF CHINA
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Problems solved by technology

[0004] Although the 16S rRNA sequences of Murexaceae species are relatively conservative, the interspecies variability is large, which makes the amplification efficiency of 16S rRNA non-specific amplification primers low, and some species cannot be amplified at all, which hinders the use of molecular techniques

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  • Muricidae mitochondria 16s rRNA gene amplification primer

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Embodiment Construction

[0007] The screening method of the amplification primer of the snail family mitochondrial 16S rRNA gene of the present invention adopts three steps: a, utilize 16S rRNA universal amplification primer to amplify the 16S rRNA sequence of the part snail family species; b, obtain from above In some 16S rRNA sequences of Mururaceae, find out the conserved sequence fragments by comparison, and synthesize multiple sets of amplification primer sequences; c, screen and amplify the mitochondrial 16S rRNA gene of Mururaceae from the multiple sets of synthetic amplification primer sequences The best pair of primers.

[0008] The present invention will be further described below in conjunction with embodiment:

[0009] 1. Sample collection: From 2005 to 2010, a total of 160 individuals of 10 genera and 25 species of Murexaceae were collected from the north and south coasts of China.

[0010] 2. DNA extraction: DNA extraction was performed on the collected samples by CTAB method.

[0011]...

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Abstract

The invention discloses a muricidae mitochondria 16S rRNA gene amplification primer, which is characterized by 16S rRNAar:5'-CGCCTGTTTATC AAAAACAT-3' and 16S rRNAbr:5'-CCGGTCTGAACTCAGATCACGT-3', AND comprises the three steps of: a, amplifying 16S rRNA sequences of part of muricidae species by 16S rRNA universal amplification primer; b, locating conserved sequence fragments in the resultant 16S rRNA sequences of part of muricidae species by means of comparison, and synthesizing the sequence fragments into multiple amplification primer sequences; and c, screening the synthesized amplification primer sequences to obtain a pair of primers having the best effect of amplifying muricidae mitochondria 16S rRNA gene. With the muricidae mitochondria 16S rRNA gene amplification primer according to the invention, 16S rRNA sequences of different muricidae species can be effectively amplified, which is significant to the research, through 16S rRNA sequences, on the classification of muricidae species and to the protection of muricidae species resource.

Description

technical field [0001] The invention relates to a primer for amplifying the 16S rRNA [6S ribosomal RNA (16small-subunit ribosomal RNA)] gene of shellfish mitochondria, in particular to an amplifying primer for the 16S rRNA gene of the snail family mitochondria. Background technique [0002] The snail family (Muricidae) belongs to Gastropoda (Gastropoda), Prosobranchia (Prosobranchia), and Neogastropoda (Neogastropoda), which are distributed in sea areas all over the world. Second major. Murex species play an important role in maintaining the ecological balance of the ocean intertidal zone, and some species can also be used as indicators for the detection of marine toxic pollutants tributyltin (TBT). Some species of the genus Rapana and Thais have delicious meat and are of great economic value. However, due to the high variability in the external shell morphology and radula structure of Mururidae, it is extremely difficult to use morphological characteristics to analyze the...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
Inventor 邹山梅李琪孔令锋
Owner OCEAN UNIV OF CHINA
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