Team:TU Darmstadt/protocols/Heat Shock Transformation

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<h2><font size="6" color="#F0F8FF" face="Arial regular"> Heat Shock Transformation </font></h2>
<h2><font size="6" color="#F0F8FF" face="Arial regular"> Heat Shock Transformation </font></h2>
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<B> Materials<br></B></font></p>
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<B> Materials<br></B>
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Ice<br>
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<p text-aligne:left style="margin-left:60px; margin-right:50px"><font size="3" color="#F0F8FF" face="Arial regular">
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Heatingbad<br>
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Pipets + steril tips<br>
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DYT-Media<br>
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LB-Agar-Plates + antibiotics<br>
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Incubator<br>
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<br>
<br>
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<B> Procedure<br></B>
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<B>Equipment<br></B>
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1. Defrost stocks of competent cells (100 µL in 1.5 ml Eppendorf tube) on ice<br>
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<div align="left" style="margin-left:60px; margin-right:50px">
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2. Add the DNA and incubate the suspension for 5 minutes on ice<br>
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<ul>
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3. Heat shock is done by incubating the cells for 1 minute at 42°C<br>
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<li class=list1>- Heating bath</li>
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4. Add 1 mL of DYT medium and incubate for 1 hour at 37°C in order to obtain antibiotic resistance<br>
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<li class=list1>- Incubator</li>
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5. Finally the cells are ready to be crossed out<br>
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</ul>
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<p text-aligne:left style="margin-left:60px; margin-right:50px"><font size="3" color="#F0F8FF" face="Arial regular">
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<br>
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<B>Chemicals & consumables<br></B>
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<div align="left" style="margin-left:60px; margin-right:50px">
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<ul>
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<li class=list1>- Ice</li>
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<li class=list1>- Pipets + steril tips</li>
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<li class=list1>- <a href="https://2013.igem.org/Team:TU_Darmstadt/materials/Media"> dYT Medium</a></li>
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<li class=list1>- LB-Agar-Plates + antibiotics</li>
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</ul>
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</div>
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</font>
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</p>
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<br>
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<p text-aligne:left style="margin-left:50px; margin-right:50px"><font size="4.5" color="#F0F8FF" face="Arial regular">
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<B> Procedure<br></B></font></p><br>
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<p text-aligne:left style="margin-left:60px; margin-right:60px"><font size="3" color="#F0F8FF" face="Arial regular">
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<div align="left" style="margin-left:30px; margin-right:50px">
 +
<ol>
 +
<li>Defrost stocks of competent cells (100 µL in 1.5 ml Eppendorf tube) on ice.</li>
 +
<li>Add the DNA and incubate the suspension for 5 minutes on ice.</li>
 +
<li>Heat shock is done by incubating the cells for 1 minute at 42°C.</li>
 +
<li>Add 1 mL of DYT medium and incubate for 1 hour at 37°C in order to obtain antibiotic resistance.</li>
 +
<li>Finally the cells are ready to be crossed out.</li>
 +
</ol>
 +
</div>
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</font></p>
<br>
<br>

Latest revision as of 00:32, 5 October 2013







Lab book | Materials | Protocols

Heat Shock Transformation

Materials


Equipment

  • - Heating bath
  • - 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.


Retrieved from "http://2013.igem.org/Team:TU_Darmstadt/protocols/Heat_Shock_Transformation"