Revision as of 17:07, 17 September 2015

Transformation of plasmids into electrocompetent cells

1. Thaw out the 50 µL cell suspension aliquot on ice for ca. 30 minutes. Concurrently, cool electroporation tubes on ice.
2. Add 100pg-1µg of plasmid DNA to the cell suspension (approximately 1-5 µL of plasmid solution).
3. Mix plasmid and cells thoroughly and continue to incubate on ice for 5-10 minutes.
4. Transfer the mixture of cells and plasmids to an electroporation tube.
5. Electroporate cells; for E.coli, this is done at 2.5 kV, 200 Ohms, 25 uF.
6. Immediately add 450-950 uL of LB broth.
7. Incubate tubes at 37^oC for 60-90 minutes in a statis incubator.
8. Plate 100 uL of cells onto agar plates with appropriate antibiotic resistance.
9. Incubate overnnight at 37^oC in a static incubator.

@@ Line 14: / Line 14: @@
 	<li>7.	Incubate tubes at 37<sup>o</sup>C for 60-90 minutes in a statis incubator.</li>
          <li>8.	Plate 100 uL of cells onto agar plates with appropriate antibiotic resistance.</li>
-	<li>8.	Incubate overnnight at 37<sup>o</sup>C in a static incubator.</li>
+	<li>9.	Incubate overnnight at 37<sup>o</sup>C in a static incubator.</li>
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