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Primer group for identifying storage book lice and application of primer group

A technology for storing book lice and book lice, which is applied in the directions of recombinant DNA technology, microbial determination/inspection, biochemical equipment and methods, etc. Easy to operate and use, low cost, accurate and reliable identification results

Active Publication Date: 2016-04-13
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The traditional identification of booklice species is mostly based on the morphological characteristics of female adults, which has the disadvantages of requiring high professional skills and taking a long time, especially the accurate identification of non-adult insects, such as eggs, larvae, and pupae, is difficult to achieve, causing quarantine work. very difficult

Method used

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  • Primer group for identifying storage book lice and application of primer group
  • Primer group for identifying storage book lice and application of primer group
  • Primer group for identifying storage book lice and application of primer group

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1. Design of specific primer pairs

[0073] The present invention designs specific primer pairs for eight common storage book lice based on the mitochondrial COI gene DNA barcode segment of ten kinds of common storage book lice, after multiple sequence comparison analysis, as shown in Table 1.

[0074] Table 1 List of 8 common storage book lice specific primers in the world

[0075]

[0076]

Embodiment 2

[0077] Example 2. Specific detection of primers

[0078] 1. Specific detection of entoF3 and entoR2

[0079] (1) Using the CTAB method to extract the genomic DNA of the single head (or multiple heads) of the storage book lice from sample 1 to sample 21 respectively.

[0080] (2) Use each genomic DNA obtained in step (1) as a template, and use entoF3 and entoR2 as primers for PCR amplification to obtain each PCR amplification product, and use ddH 2 O replaces the template as a negative control.

[0081] PCR amplification system (total volume 25μL): 2×TaqPCRMasterMix (purchased from Shenggong Bioengineering (Shanghai) Co., Ltd.) 12.5μL, ddH 2 O9.5μL, 1μL of template with a concentration of 100-300ng / μL, and 1μL of primers with a concentration of 10μM.

[0082] PCR amplification program: 94°C pre-denaturation for 3min; 94°C denaturation for 30sec, 50°C annealing for 30sec, 72°C extension for 30sec, 30 cycles; 72°C extension for 5min.

[0083] (3) Perform agarose gel electrophoresis on each ...

Embodiment 3

[0115] Example 3. Sensitivity detection of primers

[0116] 1. Sensitivity detection of entoF3 and entoR2

[0117] (1) Use CTAB method to extract the genomic DNA of sample 2 and dilute it to 0.01ng / μL, 0.1ng / μL, 1ng / μL, 5ng / μL, 10ng / μL, 25ng / μL, 50ng / μL, 75ng / μL and 100ng / μL concentration.

[0118] (2) Using the genomic DNA of each dilution obtained in step (1) as a template, and using entoF3 and entoR2 as primers for PCR amplification, each PCR amplification product is obtained, and ddH2O is used as a negative control instead of the template.

[0119] The PCR amplification system is as in Example 2.

[0120] The PCR amplification procedure is as in Example 2.

[0121] (3) Perform agarose gel electrophoresis on each PCR amplified product, and the result is as follows Picture 9 Shown.

[0122] Picture 9 Among them, M: DNA relative molecular weight standard (D2000); 1 to 9 are 0.01ng / μL, 0.1ng / μL, 1ng / μL, 5ng / μL, 10ng / μL, 25ng / μL, 50ng / μL, 75ng / μL and 100ng / μL 9 different concentratio...

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Abstract

The invention discloses a primer group for identifying storage book lice and application of the primer group. The primer group comprises at least one primer pair. The primer pairs comprise DNA (deoxyribonucleic acid) molecules which are respectively shown as (1) SEQ ID No.1, SEQ ID No.2, SEQ ID No.3, SEQ ID No.4, SEQ ID No.5, SEQ ID No.6, SEQ ID No.7 and SEQ ID No.8. The primer group and the application have the advantages that the species of the storage book lice can be quickly and efficiently identified by primers, identification results are accurate and reliable, the primer group is high in detection sensitivity, and high-throughput detection can be implemented; identification objects are free of influence of species gender, developmental stages and integrity degrees of morphological identification characters as compared with the traditional morphological identification methods; identification procedures are easy and convenient to implement and are economical and feasible.

Description

Technical field [0001] The invention relates to a set of primers for identifying storage book lice and their applications, and belongs to the field of biotechnology. Background technique [0002] Booklice, also known as paperlice and ricelice, belong to the order Psocoptera, Liposcelididae, Liposcelididae, and LiposcelisMotschμlsky1852. Booklice insects are small in size, with a body length of 0.6-1.5mm, and are widely distributed around the world; most of them live indoors and a few live outdoors. They are common pests in storage protection. This kind of pests have the characteristics of strong activity, wide feeding range, short growth cycle, diverse reproduction methods, carrying a variety of germs, strong resistance and difficulty in prevention and control, etc., which seriously harm storage goods and threaten human health. Book lice pests can spread long-distance along with stored goods. With the increasing frequency of international trade and storage of grains, the risk of...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 李志红杨倩倩
Owner CHINA AGRI UNIV
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