Team:UC Davis/Notebook/Protocols

From 2012.igem.org

(Difference between revisions)
Line 411: Line 411:
<article>
<article>
<hp>Transformations</hp>
<hp>Transformations</hp>
-
<br>We do this here
+
<p>Materials</p>
 +
•Competent Cells
 +
<br>•DNA template
 +
<br>•800 ul LB
 +
<br>•LB+Antibiotic Plates
 +
<p>Procedure</p>
 +
•Thaw competent cells on ice.
 +
<br>•Transfer 50 uL of competent cells to chilled falcon tubes.
 +
<br>•Add 1 uL of template to cells (2.5 uL if dilute).
 +
<br>•Incubate on ice for 30 minutes.
 +
<br>•Heat schock in 42 °C water bath for 90 seconds.
 +
<br>•Immediately place back onto ice for 2 minutes.
 +
<br>•Add 800 uL of LB to each tube.
 +
<br>•Incubate at 37 °C for 1 hour.
 +
<br>•Place 200 uL of the transformed cells on plates containing LB and the appropriate antibiotic.
 +
<br>•Incubate overnight at 37 °C.
</article>
</article>
   </div>
   </div>

Revision as of 22:57, 31 July 2012

Team:UC Davis - 2012.igem.org


Protocols

These are our protocols.

Registry Part Distribution Rehydration
•Add 20ul sterile H2O to desired well in distribution plate.
•Incubate at room temperature for ~10 minutes.
•Transfer resuspension to microcentrifuge tube.

Transformations

Materials

•Competent Cells
•DNA template
•800 ul LB
•LB+Antibiotic Plates

Procedure

•Thaw competent cells on ice.
•Transfer 50 uL of competent cells to chilled falcon tubes.
•Add 1 uL of template to cells (2.5 uL if dilute).
•Incubate on ice for 30 minutes.
•Heat schock in 42 °C water bath for 90 seconds.
•Immediately place back onto ice for 2 minutes.
•Add 800 uL of LB to each tube.
•Incubate at 37 °C for 1 hour.
•Place 200 uL of the transformed cells on plates containing LB and the appropriate antibiotic.
•Incubate overnight at 37 °C.

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