Vanessa cardui embryo injection
1. Collect Vanessa embryos on plant for ~3 h. This collection range has worked well for us when performing dsRNA injections; other collection intervals may be optimal for somatic or germline CRISPR/Cas9 experiments.
2. During collection, prepare the agar sheets. Bring a solution of 3% Drosophila food-grade agar to boil, then pour into 3D-printed molds. Get STL files to make 3D-printed agar molds here (files S3 and S4).
3. Collect embryos from plant, allowing them to fall onto a wide tray below.
4. Using water and a brush, line up embryos into grooves of agar sheet, as seen in video below.
1. Collect Vanessa embryos on plant for ~3 h. This collection range has worked well for us when performing dsRNA injections; other collection intervals may be optimal for somatic or germline CRISPR/Cas9 experiments.
2. During collection, prepare the agar sheets. Bring a solution of 3% Drosophila food-grade agar to boil, then pour into 3D-printed molds. Get STL files to make 3D-printed agar molds here (files S3 and S4).
3. Collect embryos from plant, allowing them to fall onto a wide tray below.
4. Using water and a brush, line up embryos into grooves of agar sheet, as seen in video below.
5. Use the back end of a brush to gently press each embryo into the agar, as seen in video below. The embryos tend to stick to the end of the needle, and we find this step helps hold the embryo in place during injection.
6. Inject embryos as seen below: