Team:TU Darmstadt/protocols/Heat Shock Transformation

From 2013.igem.org

(Difference between revisions)
Line 263: Line 263:
<li class=list1>- Ice</li>
<li class=list1>- Ice</li>
<li class=list1>- Pipets + steril tips</li>
<li class=list1>- Pipets + steril tips</li>
-
<li class=list1>- dYT-Media</li>
+
<li class=list1>- <a href="https://2013.igem.org/Team:TU_Darmstadt/materials/Media"> dYT Medium</a></li>
<li class=list1>- LB-Agar-Plates + antibiotics</li>
<li class=list1>- LB-Agar-Plates + antibiotics</li>
</ul>
</ul>

Revision as of 23:46, 4 October 2013





Heat Shock Transformation

Materials


Equipment

  • - Heatingbad
  • - Incubator


Chemicals & consumables

  • - Ice
  • - Pipets + steril tips
  • - dYT Medium
  • - LB-Agar-Plates + antibiotics


Procedure


  1. Defrost stocks of competent cells (100 µL in 1.5 ml Eppendorf tube) on ice.
  2. Add the DNA and incubate the suspension for 5 minutes on ice.
  3. Heat shock is done by incubating the cells for 1 minute at 42°C.
  4. Add 1 mL of DYT medium and incubate for 1 hour at 37°C in order to obtain antibiotic resistance.
  5. Finally the cells are ready to be crossed out.


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