Difference between revisions of "Team:Liceo Eugenio Hostos/Experiments"

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Experiments & Protocols

Protocols

Bacterial transformation:
• remove the biobriks that come in kits plates.
• Put 10 cl water HQ in the well and pump that blended leaving 10 ul of a red substance. Then put the substance in had ependorf.
• It should then be left with chemo-competent bacteria.
  
• Introducing: 100 ul of bacteria.
  
• Add 250 ul only to bacteria.
  
• 2 ul plasmido.
  
• 10 cold 0 °.
  
• 42° min 50 sec Thermo blocked.
  
• 2 min 0 °.
  
• 10 min T ° ambiente.
  
• LB+Antibiotico.
• Lb+CHLOR.
• LB+Bacteria.
• LB+Biobrick CHLOR.
• 5ml.
• 5 antibiotic to Biobrick.
• LB CHLOR.
• 10 ul min T ° environment.
• Place all the bacteria in a culture.

Purification plasmodio:

1. place 1 ml of bacteria to ependolf tubes then put them for 5 min at 14000 to the centrifuga.
2. discard liquid and leave the pellet, then we went back to step 1. Then we went back to throw the pellet. This was to get more crop.
3. place 250 ul of resuspencion solution.
4. place 250 ul of lisis.
5. solution - place 10 ul of solution - alkaline protease, wait 5min
6. place 350 ul of neutralizing solution and wait 10 min, centrifuge at 10 min and max velocidad.
7. insert the sample 600 UL listed in a column and in a tube colector.
8. remove lysate and reinsertamos column in the tube colector.
9. centrifuge at max speed by 1 min.
10. Add 750 ul of lavado.
11. solution - return to step 10.
12. we discard the excess and reinsertamos in the way.
13. column - add 250ul solution of washing.
14. centrifuge at max speed for 2 min
15. transfer the column to a tube esteril.
16. Add 80 ul of free water nucleasa.
17. centrifuge at max speed by 1 min.
18. keep the sample at - 20° C.