Transforming chemically competent cells (Inoue) protocol - source code
From OpenWetWare
#include "BioStream.h"
void main()
{
start_protocol("Transformation of chemically competent cells - Inoue");
Fluid cells = new_fluid("TB buffer cells");
Fluid dna = new_fluid("DNA");
Fluid soc = new_fluid("SOC", RT);
Plate plate = new_plate("plate made with appropriate antibiotic");
Container tube1 = new_container(STERILE_MICROFUGE_TUBE);
//1. Thaw 25 - 200 μl TB buffer cells on ice. Do not use glass tubes, which adsorb DNA.
first_step();
measure_fluid(cells, vol_range(25, 200, UL), tube1);
store_until(tube1, ON_ICE, THAW_ICE);
comment("Do not use glass tubes which adsorb DNA.");
//2. Add DNA, pipette gently to mix (keep volume of DNA less than 5% of the cell volume)
next_step();
measure_fluid(dna, tube1);
pipet(tube1);
comment("Keep volume of DNA less than 5% of the cell volume.");
//3. Incubate on ice for 30 minutes
// * Note: If you are in a rush, you can shorten this incubation time to 5-10 min.
next_step();
incubate(tube1, ON_ICE, time(30, MINS));
comment("Note: If you are in a rush, you can shorten this incubation time to 5-10 min.");
// 4. Incubate cells for 30 seconds at 42oC.
next_step();
incubate(tube1, 42, time(30, SECS));
// 5. Incubate cells on ice for 2 min.
next_step();
incubate(tube1, ON_ICE, time(2, MINS));
//6. Add 4 volumes of room temperature SOC (not critical)
next_step();
measure_prop_and_add(tube1, soc, 4);
comment("(not critical)");
// 7. Incubate for 1 hour at 37oC on shaker.
// * Note: Can also save some time here by reducing incubation to ~45 min.
// * Note: Step can be eliminated if plating on Amp plates, but not most other antibiotics
next_step();
incubate(tube1, 37, time(1, HRS), 200);
comment("Note: Can also save some time here by reducing incubation to ~45 min.");
comment("Note: Step can be eliminated if plating on Amp plates, but not most other antibiotics.");
// 8. Spread 100-300 μl onto a plate made with appropriate antibiotic.
next_step();
plate_out(plate, tube1, vol_range(100, 300, UL));
// 9. Grow overnight at 37 °C.
next_step();
incubate_plate(plate, 37, time(12, HRS));
end_protocol();
}