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Dermatophagoides farinae microsatellite marker, primer and application thereof, and acquisition method of primer

A technology of microsatellite markers and dust mites, applied in biochemical equipment and methods, measurement/testing of microorganisms, DNA/RNA fragments, etc., can solve the problems of lack of molecular markers, etc., and achieve stable PCR amplification and high polymorphism , the effect of high application value

Pending Publication Date: 2021-12-21
WANNAN MEDICAL COLLEGE
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Problems solved by technology

At present, there are few studies on the population genetics of D. farinae, mainly due to the lack of effective molecular markers

Method used

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  • Dermatophagoides farinae microsatellite marker, primer and application thereof, and acquisition method of primer
  • Dermatophagoides farinae microsatellite marker, primer and application thereof, and acquisition method of primer
  • Dermatophagoides farinae microsatellite marker, primer and application thereof, and acquisition method of primer

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Experimental program
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Embodiment

[0121] 1) The DNA of 1,000 D. farinae was extracted using the phenol-chloride method, and the extracted D. farinae genomic DNA was temporarily stored in a refrigerator at -20 degrees Celsius. After passing the test, the Illumina HiSeqTM platform was used for sequencing. The sequencing strategy was IlluminaPE150, and the total sequencing read length was 300bp.

[0122] 2) Using NCBI's existing D. farinae genome sequence as a reference gene, use the BWA software to compare the sequenced data. After correcting the BestPractices process of GATK, use the software MISA (http: / / pgrc.ipk-gatersleben.de / misa / misa.html) for microsatellite identification analysis, the software specifies specific parameters, that is, single nucleotides with more than 12 single-base repeats, 6 or more di-base repeats, and 5 three- and four-base repeats times or more, the sequence of this repeat type is a microsatellite sequence. The filter results are as follows:

[0123] A total of 266 microsatellite m...

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Abstract

The invention discloses a dermatophagoides farinae microsatellite marker. The microsatellite marker is selected from at least one of Df01, Df02, Df03, Df04, Df05, Df06, Df07, Df08, Df09, Df10, Df11 and Df12, wherein Df01 to Df12 are DNA sequences of SEQ ID NO.01 to Df12 or complementary sequences thereof in sequence, and microsatellite repetition elements of the Df01 to Df12 are (CTG)5-8, (ATG)5-7, (ATC)9-12, (GTT)6-7, (ATC)6-8, (TG)6-9, (ATG)5-9, (GT)7-10, (ATG)4-5, (CAA)5-7, (TTG)6-9 and (CA)7-8 The microsatellite marker and the primer thereof provided by the invention have the characteristics of stable PCR amplification, high polymorphism and the like, and the primer can be used for genetic diversity and genetic structure analysis of dermatophagoides farinae and has relatively high application value.

Description

technical field [0001] The invention relates to the technical field of molecular biology DNA markers, in particular to a dust mite microsatellite marker, primers and applications thereof, and a primer acquisition method. Background technique [0002] Dust mites, belonging to the family Acaridae, have a wide range of habitats and often breed in house dust, air-conditioning dust screens, flour, biscuit powder, animal feed, Chinese medicinal materials, and waste cotton. Domestically, it is mainly distributed in the southern and central regions of China. Dust mites are ubiquitous in the indoor environment where human beings live and work, and their excrement, metabolites, shed shells and dead mites are strong allergens, which can cause mite allergic diseases, mite dermatitis and Mites in humans etc. [0003] At present, there is no specific drug for mite allergic diseases caused by dust mite allergens. Effective control of the number of dust mites in the environment has a posi...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6811C12N15/11
CPCC12Q1/6888C12Q1/6811C12Q2600/156C12Q2600/124C12Q2531/113C12Q2535/122C12Q2565/125C12Q2537/165
Inventor 孙恩涛刘艺方惟希李咏琪王慧慧李梦楠王铎黄庆宇
Owner WANNAN MEDICAL COLLEGE
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