11. Use tube immediately for electroporation or store in a À80
C
freezer (see Note 15).
12. Before electroporation, thaw an Eppendorf tube on ice if
necessary.
13. Centrifuge at 5500 rpm (4100 Â g) for 5 min at 4
C, discard
the supernatant and resuspend the bacteria in 1 mL ice-cold
PBS pH 7.4 (see Note 14).
14. Centrifuge tubes again at 5500 rpm (4100 Â g) for 5 min at
4
C, discard the supernatant and resuspend the bacteria in
5 mL ice-cold PBS pH 7.4 in a 15 mL prechilled tube (see Note
14).
15. Add 4 μL of YOPRO solution to the flask.
3.3 Flow-Through
Electroporation
Prepare the Franz diffusion cell bottom receiver chamber (Fig. 3):
1. Connect the water jacket inlet and outlet to a reservoir with
ice-cold water, and use a pump to flow water through the jacket
to precool the diffusion cell.
Fig. 3 Schematic of the flow-through electroporation device. Dimensions are not to scale. Inset shows relative
sizes of the E. coli in blue and the openings of the gold microtubes (reprinted with permission from ref. 11.
Copyright 2016, American Chemical Society)
48
Juliette Experton et al.
C
freezer (see Note 15).
12. Before electroporation, thaw an Eppendorf tube on ice if
necessary.
13. Centrifuge at 5500 rpm (4100 Â g) for 5 min at 4
C, discard
the supernatant and resuspend the bacteria in 1 mL ice-cold
PBS pH 7.4 (see Note 14).
14. Centrifuge tubes again at 5500 rpm (4100 Â g) for 5 min at
4
C, discard the supernatant and resuspend the bacteria in
5 mL ice-cold PBS pH 7.4 in a 15 mL prechilled tube (see Note
14).
15. Add 4 μL of YOPRO solution to the flask.
3.3 Flow-Through
Electroporation
Prepare the Franz diffusion cell bottom receiver chamber (Fig. 3):
1. Connect the water jacket inlet and outlet to a reservoir with
ice-cold water, and use a pump to flow water through the jacket
to precool the diffusion cell.
Fig. 3 Schematic of the flow-through electroporation device. Dimensions are not to scale. Inset shows relative
sizes of the E. coli in blue and the openings of the gold microtubes (reprinted with permission from ref. 11.
Copyright 2016, American Chemical Society)
48
Juliette Experton et al.