Team:LIKA-CESAR-Brasil/TransformationProtocol

From 2014.igem.org

(Difference between revisions)
Line 302: Line 302:
         <li><a href="https://2014.igem.org/Team:LIKA-CESAR-Brasil/Safety">SAFETY</a></li>
         <li><a href="https://2014.igem.org/Team:LIKA-CESAR-Brasil/Safety">SAFETY</a></li>
         <li><a href="https://2014.igem.org/Team:LIKA-CESAR-Brasil/Atribuitons">ATRIBUITIONS</a></li>
         <li><a href="https://2014.igem.org/Team:LIKA-CESAR-Brasil/Atribuitons">ATRIBUITIONS</a></li>
 +
        <li class="igem-logo"><a href="https://2014.igem.org/"></a></li>
             </ul>
             </ul>
           </div><!--/.nav-collapse -->
           </div><!--/.nav-collapse -->

Revision as of 02:08, 18 October 2014

LIKA | CESAR

NOTEBOOK

Transformation protocol

  • 1) Transform cells with 50 l of 1 µl plasmid.
  • 2) Dilute plasmid NEB buffer.
  • 3) Incubate the mixture of cells with the resistance plasmid for 30 minutes on ice.
  • 4) After 30 minutes, bring the sample to heat shock in a water bath at 42 ° C for 60 seconds, after adding 200 l SOC medium and incubate at 37 ° C for 2 hours. (Procedures performed in 2 ml tubes).
  • 5) Tagging cells transformed with plasmid resistance on solid LB containing antibiotics (chloramphenicol).
  • 6) 250 L of cell solution was plated on solid medium for 16 hours in an oven at 37 ° C microbiological.
  • 7) After the 16 hours incubation the counting antibiotic-resistant colonies was made (observed the efficiency of transformation).
  • 8) Calculation of transformation efficiency: Efficiency of transformation (dilution factor = 15) x colony count x 105 / μgDNA.
Marca LIKA Marca CESAR