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Team:TU Darmstadt/Notebook/Methods/Heat shock transformation
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| - | <!--TYPO3SEARCH_begin--><div id="c101" class="csc-default"><div class="csc-header csc-header-n1"><h1 class="csc-firstHeader">Heat shock transformation</h1></div><div><p><b>Equipment:</b></p></div><div><p>- Heating bath</p></div><div><p>- Incubator</p></div><div></div><div><p><b>Chemicals & consumables:</b></p></div><div><p>- Ice</p></div><div><p>- Pipets + steril tips</p></div><div><p>- LB medium</p></div><div><p>- LB-Agar-Plates + antibiotics</p></div><div></div><div><p><b>Procedure:</b></p></div><div></div><div><p>Defrost stocks of competent cells (100 µL in 1.5 ml Eppendorf tube) on ice.</p></div><div><p>Add the DNA and incubate the suspension for 15 minutes on ice.</p></div><div><p>Heat shock is done by incubating the cells for 45 seconds at 42°C. Put samples back on ice for 2 min.</p></div><div><p>Add 1 mL of LB medium and incubate for 1 hour at 37°C in order to obtain antibiotic resistance.</p></div><div><p>It might be useful to spin down cells at 5000 rpm for 5 min. Resuspend pellet in 100 µL LB.</p></div><div><p>Finally the cells are ready to be spread out. </p></div></div><!--TYPO3SEARCH_end--> | + | <!--TYPO3SEARCH_begin--><div id="c101" class="csc-default"><div class="csc-header csc-header-n1"><h1 class="csc-firstHeader">Heat shock transformation</h1></div><div><p><b>Equipment:</b></p></div><div><p>- Heating bath</p></div><div><p>- Incubator</p></div><div></div><div><p><b>Chemicals & consumables:</b></p></div><div><p>- Ice</p></div><div><p>- Pipets + steril tips</p></div><div><p>- LB medium</p></div><div><p>- LB-Agar-Plates + antibiotics</p></div><div></div><div><p><b>Procedure:</b></p></div><div></div><div><p>- Defrost stocks of competent cells (100 µL in 1.5 ml Eppendorf tube) on ice.</p></div><div><p>Add the DNA and incubate the suspension for 15 minutes on ice.</p></div><div><p>Heat shock is done by incubating the cells for 45 seconds at 42°C. Put samples back on ice for 2 min.</p></div><div><p>- Add 1 mL of LB medium and incubate for 1 hour at 37°C in order to obtain antibiotic resistance.</p></div><div><p>- It might be useful to spin down cells at 5000 rpm for 5 min. Resuspend pellet in 100 µL LB.</p></div><div><p>- Finally the cells are ready to be spread out. </p></div></div><!--TYPO3SEARCH_end--> |
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Revision as of 18:44, 17 October 2014
Heat shock transformation
Equipment:
- Heating bath
- Incubator
Chemicals & consumables:
- Ice
- Pipets + steril tips
- LB medium
- LB-Agar-Plates + antibiotics
Procedure:
- Defrost stocks of competent cells (100 µL in 1.5 ml Eppendorf tube) on ice.
Add the DNA and incubate the suspension for 15 minutes on ice.
Heat shock is done by incubating the cells for 45 seconds at 42°C. Put samples back on ice for 2 min.
- Add 1 mL of LB medium and incubate for 1 hour at 37°C in order to obtain antibiotic resistance.
- It might be useful to spin down cells at 5000 rpm for 5 min. Resuspend pellet in 100 µL LB.
- Finally the cells are ready to be spread out.
