Size selective DNA precipitation protocol - source code - Revision history

Vaishnavi Ananth at 07:20, 27 April 2010

2010-04-27T07:20:01Z

←Older revision		Revision as of 07:20, 27 April 2010
Line 38:		Line 38:
	next_step();		next_step();
	measure_fluid(buffer, eppendorf);		measure_fluid(buffer, eppendorf);
-	~~comment~~("Add appropriate volume of buffer.");	+	to_do("Add appropriate volume of buffer.");
	dissolve(eppendorf);		dissolve(eppendorf);

Vaishnavi Ananth at 05:53, 20 November 2009

2009-11-20T05:53:31Z

←Older revision		Revision as of 05:53, 20 November 2009
Line 7:		Line 7:

	Fluid dna = new_fluid("DNA sample", "DNA to be separated (e.g. PCR reaction mixture)");		Fluid dna = new_fluid("DNA sample", "DNA to be separated (e.g. PCR reaction mixture)");
-	Fluid peg = new_fluid("PEG/MgCl2", "30% (w/v) PEG 8000/30 mM MgCl2 (concentration of PEG 8000	+	Fluid peg = new_fluid("PEG/MgCl2", "30% (w/v) PEG 8000/30 mM MgCl2 (concentration of PEG 8000 can be varied to shift the size of the percipitated DNA. The concentration used here will remove DNA fragments with less than 300bp)");
-	can be varied to shift the size of the percipitated DNA.	+
-	The concentration used here will remove DNA fragments with	+
-	less than 300bp)");	+
	Fluid te = new_fluid("TE buffer", "10 mM TRIS-HCl, 1 mM EDTA, pH 8.0");		Fluid te = new_fluid("TE buffer", "10 mM TRIS-HCl, 1 mM EDTA, pH 8.0");
	Fluid buffer = new_fluid("buffer of choice");		Fluid buffer = new_fluid("buffer of choice");

Vaishnavi Ananth at 09:50, 19 November 2009

2009-11-19T09:50:04Z

←Older revision		Revision as of 09:50, 19 November 2009
Line 1:		Line 1:
	<code><pre>		<code><pre>
-	#include "~~BioStream~~.h"	+	#include "BioCoder.h"

	void main()		void main()

Vaishnavi Ananth: New page:

 #include "BioStream.h"  void main() { 	start_protocol("Size specific DNA precipitation");  	Fluid dna = new_fluid("DNA sample", "DNA to be separated (e.g. PCR reaction mixture)...


2009-10-20T10:21:11Z
New page: <code><pre> #include "BioStream.h"  void main() { 	start_protocol("Size specific DNA precipitation");  	Fluid dna = new_fluid("DNA sample", "DNA to be separated (e.g. PCR reaction mixture)...
New page
<code><pre>

#include "BioStream.h"



void main()

{

	start_protocol("Size specific DNA precipitation");



	Fluid dna = new_fluid("DNA sample", "DNA to be separated (e.g. PCR reaction mixture)");

	Fluid peg = new_fluid("PEG/MgCl2", "30% (w/v) PEG 8000/30 mM MgCl2 (concentration of PEG 8000 

                                                 can be varied to shift the size of the percipitated DNA.

                                                 The concentration used here will remove DNA fragments with 

                                                 less than 300bp)");

	Fluid te = new_fluid("TE buffer", "10 mM TRIS-HCl, 1 mM EDTA, pH 8.0");

	Fluid buffer = new_fluid("buffer of choice");



	Container eppendorf = new_container(EPPENDORF);



	//* Mix 50 μL of sample with 150 µL of TE

	first_step();

	measure_fluid(dna, vol(50, UL), eppendorf);

	measure_fluid(te, vol(150, UL), eppendorf);

	tap(eppendorf);



	//* Add 100 µL of PEG/MgCl2

	next_step();

	measure_fluid(peg, vol(10, UL), eppendorf);



	//* Vortex

	next_step();

	vortex(eppendorf);



	//* Centrifuge 15 min at 10.000 rcf at roomtemperature

	next_step();

	centrifuge_pellet(eppendorf, speed(10000, G), RT, time(15, MINS));



	//* Carefully remove supernatant not to disturb the pellet, which will be invisible

	next_step();

	comment("Carefully remove supernatant not to disturb the pellet, which will be invisible.");



	//* Dissolve the pellet in a appropriate amount of buffer of choice 

	next_step();

	measure_fluid(buffer, eppendorf);

	comment("Add appropriate volume of buffer.");

	dissolve(eppendorf);



	end_protocol();

}

</pre></code>