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Team:Cambridge/Protocols/Transformation of E.coli by Electroporation
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| - | == | + | ==Transformation of E.coli by Electroporation== |
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| + | This protocol describes how to transform cells prepared for electroporation using concentrated DNA and an electroporation cuvette. | ||
===Theory=== | ===Theory=== | ||
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| + | Although imaging data was not available at the time of Weaver and Chizmadzhev's review of the "Theory of Electroporation" they discuss detailed indirect evidence and mathematical models which strongly suggest that electroporation causes aqueous pores in lipid membranes and that molecules can enter by electric drift. A charged molecule such as DNA is thus very likely to be affected by the electric field applied to cells in electroporation. | ||
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| + | J. C. Weaver and Y. A. Chizmadzhev."Theory of electroporation: A review " Biochemistry and Bioenergetics. 41. (1996) 135-160 | ||
===Practice=== | ===Practice=== | ||
How to do it in the lab | How to do it in the lab | ||
| - | + | *Step 1 Prepare Competent Cells as per the Competent Cells for Electroporation Protocol | |
| - | + | *Step 2 Dilute your superconcentrated DNA (Ingenio Electroporation kits recommend the DNA displays an absorbance at 280nm of 1.8 - 2.0 | |
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===Safety=== | ===Safety=== | ||
Revision as of 19:21, 30 August 2011
Transformation of E.coli by Electroporation
This protocol describes how to transform cells prepared for electroporation using concentrated DNA and an electroporation cuvette.
Theory
Although imaging data was not available at the time of Weaver and Chizmadzhev's review of the "Theory of Electroporation" they discuss detailed indirect evidence and mathematical models which strongly suggest that electroporation causes aqueous pores in lipid membranes and that molecules can enter by electric drift. A charged molecule such as DNA is thus very likely to be affected by the electric field applied to cells in electroporation.
J. C. Weaver and Y. A. Chizmadzhev."Theory of electroporation: A review " Biochemistry and Bioenergetics. 41. (1996) 135-160
Practice
How to do it in the lab
- Step 1 Prepare Competent Cells as per the Competent Cells for Electroporation Protocol
- Step 2 Dilute your superconcentrated DNA (Ingenio Electroporation kits recommend the DNA displays an absorbance at 280nm of 1.8 - 2.0
Safety
The safety implication of the procedure.
