Transforming chemically competent cells (Inoue) protocol - source code

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(New page: <code> <pre> #include "BioStream.h" void main() { start_protocol("Transformation of chemically competent cells - Inoue"); Fluid cells = new_fluid("TB buffer cells"); Fluid dna = n...)
Current revision (01:33, 20 November 2009) (view source)
 
Line 1: Line 1:
<code>
<code>
     <pre>
     <pre>
-
#include "BioStream.h"
+
#include "BioCoder.h"
void main()
void main()
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Fluid cells = new_fluid("TB buffer cells");
Fluid cells = new_fluid("TB buffer cells");
-
Fluid dna = new_fluid("DNA");
+
Fluid dna = new_fluid("DNA",vol(20, UL));
Fluid soc = new_fluid("SOC", RT);
Fluid soc = new_fluid("SOC", RT);
Plate plate = new_plate("plate made with appropriate antibiotic");
Plate plate = new_plate("plate made with appropriate antibiotic");
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//6. Add 4 volumes of room temperature SOC (not critical)
//6. Add 4 volumes of room temperature SOC (not critical)
next_step();
next_step();
-
measure_prop_and_add(tube1, soc, 4);
+
measure_prop(tube1, soc, 4);
comment("(not critical)");
comment("(not critical)");

Current revision

#include "BioCoder.h"

void main()
{
	start_protocol("Transformation of chemically competent cells - Inoue");

	Fluid cells = new_fluid("TB buffer cells");
	Fluid dna = new_fluid("DNA",vol(20, UL));
	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(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();
}
    

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