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<div class="col-xs-12 col-sm-12 col-md-12 col-lg-12"> | <div class="col-xs-12 col-sm-12 col-md-12 col-lg-12"> | ||
− | <h2>Requirements for each sample</h2> | + | <h2>Requirements for each sample:</h2> |
<ul class="points"> | <ul class="points"> | ||
<li><span><i class="fa fa-caret-right" aria-hidden="true"></i></span> 1 tube of thawed chemically competent cells (keep covered in ice at all times). </li> | <li><span><i class="fa fa-caret-right" aria-hidden="true"></i></span> 1 tube of thawed chemically competent cells (keep covered in ice at all times). </li> | ||
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</li> | </li> | ||
</ul> | </ul> | ||
+ | </div> | ||
+ | </div> | ||
+ | <hr class="mainline"> | ||
+ | <div class="row"> | ||
+ | <div class="col-xs-12 col-sm-12 col-md-12 col-lg-12"> | ||
+ | <h2>Procedure:</h2> | ||
+ | <ul class="points"> | ||
+ | <li><span><i class="fa fa-caret-right" aria-hidden="true"></i></span> Add DNA to cells</li> | ||
+ | <li><span><i class="fa fa-caret-right" aria-hidden="true"></i></span> Incubate on ice for 0-20 minutes</li> | ||
+ | <li><span><i class="fa fa-caret-right" aria-hidden="true"></i></span> Heat shock at 42 °C for 30 seconds</li> | ||
+ | <li><span><i class="fa fa-caret-right" aria-hidden="true"></i></span> Incubate on ice for 0-2 minutes | ||
+ | <li><span><i class="fa fa-caret-right" aria-hidden="true"></i></span> Add SOC</li> | ||
+ | <li><span><i class="fa fa-caret-right" aria-hidden="true"></i></span> Incubate on rotator in 37 °C incubator for 0-60 minutes | ||
+ | </li> | ||
+ | <li><span><i class="fa fa-caret-right" aria-hidden="true"></i></span> Plate 50-250 uL | ||
+ | </li> | ||
+ | </ul> | ||
+ | </div> | ||
+ | </div> | ||
+ | </section> | ||
+ | </body> | ||
+ | </html> |
Revision as of 18:27, 1 December 2016
Transformation
Requirements for each sample:
- 1 tube of thawed chemically competent cells (keep covered in ice at all times).
- 1-10 uL ligation product cloned in plasmid.
- 250 uL SOC
- 1 LB plate with appropriate antibiotics
Procedure:
- Add DNA to cells
- Incubate on ice for 0-20 minutes
- Heat shock at 42 °C for 30 seconds
- Incubate on ice for 0-2 minutes
- Add SOC
- Incubate on rotator in 37 °C incubator for 0-60 minutes
- Plate 50-250 uL