Small Molecule Nematicides Screening Assay: A Medium Throughput Method to Screen Potential Nematicides Against Ditylenchus dipsaci

Abstract

Source: Cammalleri, S. R., et al. Culturing and Screening the Plant Parasitic Nematode Ditylenchus dipsaci. J. Vis. Exp. (2022).

In this video, we demonstrate a screening assay to identify the efficacy of small-molecule nematicides against nematodes. Nematicides are considered effective if they cause a decrease in the number of mobile nematodes in the sample after exposure.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board

1. In vitro small molecule screen

1. Prepare the assay plates following the steps below.
   1. Pour autoclaved distilled water into a sterile trough and, using a multichannel pipette, dispense 40 µL of distilled water from the trough into each well of a flat-bottom 96-well plate.
2. Prepare pinning tool and add the chemicals.
   1. Set up pinner trays (see Table of Materials) near a Bunsen burner flame on a lab....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
50mL beaker	Pyrex	CLS100050
96 well plate	Sarstedt	83.3924
Aluminium foil	Alcan Plus
Dissecting scope	Leica	Leica MZ75
DMSO	Sigma-Aldrich	472301-500ML
Glass slide	MAGNA	60-1200
Lint-Free Blotting Paper	V&P Scientific	VP 522-100
Low retention pipette tips (200uL)	LABFORCE	1159M44
Multichannel pipette	Eppendorf research plus	3125000036
NaOH	Sigma-Aldrich	S8045-500G
Parafilm	Bemis	PM-996
Pin cleaning solutions	V&P Scientific	VP110A
Pinner	V&P Scientific	VP381N
Pinner rinse trays	V&P Scientific	VP 421
Reagent reservoir with lid for multichannel pipettes	Sigma-Aldrich	BR703459
Shaking incubator	New Brunswick Scientific	I26