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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> | + | <!--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 18:46, 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.
