Agarose-coated Petri plates containing FASW-T
MATERIALS
Reagents
Agarose, high-quality (e.g., UltraPure Agarose; Invitrogen)
Equipment
Humidified container at 4°C (optional; see Step 5)
Microwave
Petri dishes, plastic
Use 60- × 15-mm dishes for dechorionation and for growing a few hundred embryos. Use 100- × 15-mm dishes for microinjection or if handling more embryos.
METHOD
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1. Arrange the desired number of Petri plates in rows on the benchtop, and remove the lids.
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2. Use the microwave to melt a solution of 0.9%-1.0% agarose in FASW-T.
Although the percentage of agarose is not critical, it is important to minimize evaporation of H2O while heating. We have found that overboiling agarose has an adverse effect on Ciona embryo development, even though the agarose is submerged in FASW-T.
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3. Pour the melted agarose to fill the first plate. Pour agarose from the first plate into the second plate to form a thin layer of agarose on the bottom of the second plate.
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4. Continue to pour agarose from the first plate into the remaining plates until all plates contain a thin layer of agarose. Discard any unused agarose.
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5. After the agarose has solidified, add enough FASW-T to cover the agarose, and replace the lids.
The plates can be stored in a humidified container for up to 1 mo at 4°C.
