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Pretreatment method for plant nematodes for detecting MALDI-TOF MS

A technology for plant nematodes and nematodes, which is applied in the field of mass spectrometry analysis, can solve the problems affecting the detection of plant nematodes, poor stability of results, and many background signals, and achieves the effects of stable results, easy operation, and low background signals.

Inactive Publication Date: 2015-05-06
SHENZHEN AUDAQUE DATA TECH
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Problems solved by technology

However, the pretreatment method for plant nematodes before detection is difficult to operate, resulting in poor stability of the obtained results, many and mixed background signals, and low sensitivity, which affects the detection of plant nematodes

Method used

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  • Pretreatment method for plant nematodes for detecting MALDI-TOF MS

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Embodiment 1

[0040] 1. Cleaning of root-knot nematodes

[0041] Transfer the single root-knot nematode obtained from the plant to a 1.5mL centrifuge tube (Eppendorf), vortex for 5min, centrifuge at 4000rpm for 10min, remove the supernatant; add 1mL of sterilized water, vortex for 5min, centrifuge at 4000rpm for 10min, remove Supernatant: add 1mL sterilized water, vortex for 5min, centrifuge at 4000rpm for 10min, remove supernatant, add 1mL sterilized water, set aside.

[0042] 2. Pre-treatment of single root-knot nematode

[0043] (1) On the sterilized glass slide, drop 5 μ l extract (the mixed solution of formic acid, acetonitrile and sterilized water, the volume concentration of formic acid in this mixed solution is 35%, the volume concentration of acetonitrile is 50%, the balance for sterilized water).

[0044] (2) Pick a cleaned root-knot nematode with a picking needle (the picking needle can be an insect needle) under a stereomicroscope, transfer it to the extract on the glass slide...

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Abstract

The invention discloses a pretreatment method for plant nematodes for detecting MALDI-TOF MS. The pretreatment method comprises the following steps: (1) dropwise adding an extracting liquid on sterilized slide glass, wherein the dropwise addition amount of the extracting liquid is 5-8mu l per plant nematode; (2) transferring the cleaned plant nematodes into the extracting liquid through a stereoscopic microscope, and crushing the plant nematodes, wherein the extracting liquid is used for extracting the protein of the plant nematodes and hydryolyzing the protein into polypeptide; (3) with the stereoscopic microscope, pipetting the extracting liquid in the step (2) and the crushed plant nematode through a pipette, dropwise adding the extracting liquid onto a sample plate, and air-drying; and (4) adding a matrix solution on the air-dried sample plate so as to enable the polypeptide to be dispersed on the matrix solution, and air-drying to obtain the sample for detecting MALDI-TOF MS, wherein the addition amount of the matrix solution is 1-1.5mu l per plant nematode. The method is easy to operate, good in result repeatability, low in background signal and high in sensitivity.

Description

technical field [0001] The invention belongs to the technical field of mass spectrometry, and in particular relates to a MALDI-TOF MS (abbreviation for Matrix-assisted laser desorption / ionization time-of-flight mass spectrometry, Chinese full name is matrix-assisted laser desorption ionization time-of-flight mass spectrometry) detection Plant nematode pretreatment method. Background technique [0002] The MALDI-TOF MS instrument is mainly composed of two parts: matrix-assisted laser desorption ionization source (MALDI) and time-of-flight mass analyzer (TOF). The principle of MALDI is to irradiate the co-crystal film formed by the sample and the matrix with laser light. The matrix absorbs energy from the laser and transmits it to the biomolecules. During the ionization process, the protons are transferred to the biomolecules or protons are obtained from the biomolecules, so that the biomolecules are ionized. process. It is a soft ionization technique suitable for the determ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N27/64
Inventor 龙海李芳荣李一农凌杏园冯建军郑耘谢泳桂刘亮山
Owner SHENZHEN AUDAQUE DATA TECH
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