Vaishnavi Ananth: New page:

  #include "BioCoder.h"  void main() { 	start_protocol("Mouse tissue lysis for genotyping");  	Fluid taq = new_fluid("Taq DNA polymerase 10X buffer with MgCl₂", "dilu...


Vaishnavi Ananth — Mon, 22 Mar 2010 11:36:24 GMT
New page: <code> <pre> #include "BioCoder.h"  void main() { 	start_protocol("Mouse tissue lysis for genotyping");  	Fluid taq = new_fluid("Taq DNA polymerase 10X buffer with MgCl<sub>2</sub>", "dilu...
New page
<code>

<pre>

#include "BioCoder.h"



void main()

{

	start_protocol("Mouse tissue lysis for genotyping");



	Fluid taq = new_fluid("Taq DNA polymerase 10X buffer with MgCl<sub>2</sub>", "diluted 1:10 in ddH<sub>2</sub>");

	Fluid proteinasek = new_fluid("proteinase K", "20 mg/ml");



	Solid tissue = new_solid("tail or tissue chunk");

	

	Container tube1 = new_container(STERILE_MICROFUGE_TUBE, tissue);

	Container tube2 = new_container(STERILE_MICROFUGE_TUBE);



	//tissue lysis to release DNA

	//per tail or tissue chunk add (tissue degradation is sped up if tissue/tail is cut into smaller bits):

         //* 100 µl Taq DNA polymerase 10x Reaction buffer with MgCl2 (e.g. Promega #M1883) diluted 1:10 in ddH2O

         //* +2 µl proteinase K (20 mg/ml) 

	first_step("Tissue lysis to release DNA");

	measure_fluid(taq, vol(100, UL), tube1);

	measure_fluid(proteinasek, vol(2, UL), tube1);



	//* incubate for 3h to o/n at 50°C at 300 rpm 

	next_step();

	incubate(tube1, 50, time_range(3, OVERNIGHT, HRS), 300);

	comment("proteinase K is stable over a broad pH range (4.0 to 12.5, optimum pH 8.0) and is also stable over the temperature range of 25 to 65°C.");

    

	//proteinase K inactivation

         //* 95°C for 10-20 min at 300 rpm

         //* centrifuge at 13000 rpm for 3 min and transfer supernatant (80 µl) into new eppi 

	next_step("Proteinase K inactivation");

	incubate(tube1, 95, time_range(10, 20, MINS), 300);

	centrifuge_phases_top(tube1, speed(13000, RPM), RT, time(3, MINS), vol(80, UL), tube2);

	name_sample(tube2, "sample");

	

	//storage before PCR

	//* 4°C short term, -20°C long term 

	next_step("Storage before PCR");

	use_or_store(tube2, time(1, HRS), 4, -20);



	end_protocol();

}

	

</pre>

</code>

Mouse tissue lysis for genotyping protocol - source code - Revision history

Vaishnavi Ananth: New page: `#include "BioCoder.h" void main() { start_protocol("Mouse tissue lysis for genotyping"); Fluid taq = new_fluid("Taq DNA polymerase 10X buffer with MgCl₂", "dilu...`

Mouse tissue lysis for genotyping protocol - source code - Revision history

Vaishnavi Ananth: New page: #include "BioCoder.h" void main() { start_protocol("Mouse tissue lysis for genotyping"); Fluid taq = new_fluid("Taq DNA polymerase 10X buffer with MgCl2", "dilu...

Vaishnavi Ananth: New page: `#include "BioCoder.h" void main() { start_protocol("Mouse tissue lysis for genotyping"); Fluid taq = new_fluid("Taq DNA polymerase 10X buffer with MgCl₂", "dilu...`