Transformation

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<b>Procedure</b>
<b>Procedure</b>
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Preparation: competent cells
+
Preparation: [[competent cells]]
For each plasmid:
For each plasmid:
* take competent cells from -80C freezer
* take competent cells from -80C freezer
-
* leave on ice for 10min
+
* thaw on ice for 10min
* add 2uL DNA to competent cell (label)
* add 2uL DNA to competent cell (label)
* mix with pipette
* mix with pipette
-
* sit for 30min
+
* sit on ice for 30min
* get a beaker of exactly 42C water from hot water tap
* get a beaker of exactly 42C water from hot water tap
* submerge tubes in hot water bath for exactly 90s
* submerge tubes in hot water bath for exactly 90s
* sit on ice for 5min
* sit on ice for 5min
* add 1mL LB
* add 1mL LB
-
* leave in incubator for 1h
+
* incubate for 1h
-
* spread 500uL on Petri plate labelled with appropriate antibiotics. (Dip a glass rod with ethanol and flame it to spread cells in plates)
+
* spread 500uL on Petri plate labelled with appropriate antibiotics
-
* wait for plates to dry
+
* Dip a glass rod with ethanol and flame it to spread cells in plates
-
* leave in incubator overnig
+
* wait for plate to dry
 +
* incubate overnight
 +
 
 +
[[University_of_Toronto_2006| Home]] | [[Lab Training Protocol]]

Latest revision as of 16:45, 28 June 2006

iGEM 2006 Lab Training Session I: Transformation

Key words: [http://en.wikipedia.org/wiki/Transformation_%28genetics%29 transformation], [http://en.wikipedia.org/wiki/Competent_cells competent cells]

Procedure

Preparation: competent cells

For each plasmid:

  • take competent cells from -80C freezer
  • thaw on ice for 10min
  • add 2uL DNA to competent cell (label)
  • mix with pipette
  • sit on ice for 30min
  • get a beaker of exactly 42C water from hot water tap
  • submerge tubes in hot water bath for exactly 90s
  • sit on ice for 5min
  • add 1mL LB
  • incubate for 1h
  • spread 500uL on Petri plate labelled with appropriate antibiotics
  • Dip a glass rod with ethanol and flame it to spread cells in plates
  • wait for plate to dry
  • incubate overnight

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