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Nibea japonica microsatellite DNA markers

A technology of DNA markers and microsatellites, applied in the direction of recombinant DNA technology, DNA/RNA fragments, microbial measurement/inspection, etc., can solve the problem of not providing enough genetic information and achieve good repeatability

Active Publication Date: 2015-06-24
MARINE FISHERIES RES INST OF ZHEJIANG
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Problems solved by technology

However, the above genetic markers do not provide sufficient genetic information

Method used

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  • Nibea japonica microsatellite DNA markers
  • Nibea japonica microsatellite DNA markers
  • Nibea japonica microsatellite DNA markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0035] 1. Construction of Japanese microsatellite DNA plasmid library:

[0036] 1.1 Genome extraction: extract the Japanese DNA according to the method of Aljanabi and Martinez et al. Yellow croaker genomic DNA, DNA samples were detected by electrophoresis on 0.8% agarose gel.

[0037] 1. 25'-anchored PCR amplification

[0038] The following methods refer to Tang S J, Liu Z Z, Tang W Q, et al. A simple method for isolation of microsatellites from the mudskipper (Boleophthalmus pectinirostris) without constructing a genomic library [J]. Conserv Genet, 2009, 10:1957–1959.

[0039]KKVRVRV(CT)10, KKVRVRV(AG)10, KKHBHBH(AG)10, KKVRVRV(TG)10, KKHRHRH(TG)10, GGCC(AC)8, GCGC(AG)8, GCGC(AC)8 were used as primers , using the genomic DNA extracted in step 1.1 as a template for PCR amplification, the 15 μL reaction system is as follows, including 1×EX Taq buffer, 0.2mM dNTPs, 0.2μM primers, 40ng DNA template, 1U Taq DNA polymerase (purchased from TaKaRa Company ), and the rest were mad...

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Abstract

The invention relates to the technology of molecular markers, and particularly relates to nibea japonica microsatellite DNA markers. The nibea japonica microsatellite DNA markers have microsatellite numbers of NJB01, NJB04, NJB06, NJB15 and NJB35, and have nucleotide sequences as shown in SEQ ID NO:1-5 sequentially. The six new microsatellite sites of nibea japonica as well as primer sequence and amplification method for amplifying the six microsatellite sites can be applied to studies on genetic diversity and paternity identification of different groups of nibea japonica. The nibea japonica microsatellite DNA markers have good repeatability, and are reliable and effective molecular markers.

Description

technical field [0001] The invention relates to a molecular marker technology, in particular to a microsatellite DNA marker for Japanese yellow croaker. Background technique [0002] Microsatellite DNA, also known as short tandem repeats (STR), simple repeats (SSR), simple sequence length polymorphism (SSLP), refers to the tandem repeats of 1-6 nucleotides in the genome. Nucleotide sequence; among them, the most common are dinucleotide repeats, such as ((AC / TG)n and (AG / TC)n. Different organisms show polymorphism due to the number of repetitions of the repeat motif Studies have shown that it may be the "strand slippage" phenomenon in the DNA replication process that causes the high polymorphic information content (PIC) of microsatellite DNA. The high mutation and co-dominance of microsatellite DNA Expression and its ubiquity in eukaryotic genomes make it a superior molecular marker for the analysis of population genetic diversity, identification of kinship, genome mapping a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 柴学军胡则辉王跃斌朱云海
Owner MARINE FISHERIES RES INST OF ZHEJIANG
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