Small Molecule Nematicides Screening Assay: A Medium Throughput Method to Screen Potential Nematicides Against Ditylenchus dipsaci
Published: April 30, 2023
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 Prepare the assay plates following the steps below. 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. Pre…
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The authors have nothing to disclose.
Materials
| 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 |
