Toronto/Lab Protocols/Transformation
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# Leave the plate with lid half open in the incubator to dry. Rotate the plate 180° every 10 minutes to ensure even spread of bacteria. | # Leave the plate with lid half open in the incubator to dry. Rotate the plate 180° every 10 minutes to ensure even spread of bacteria. | ||
# Once dry, turn the plates upside down and incubate overnight. | # Once dry, turn the plates upside down and incubate overnight. | ||
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Jump to [http://igem.skule.ca/lab/protocols/transformation.htm BlueGenes] | Jump to [http://igem.skule.ca/lab/protocols/transformation.htm BlueGenes] | ||
Latest revision as of 03:31, 27 October 2007
Transformations
4 hours
Transformation is a genetic alteration of a cell resulting from the introduction, uptake and expression of foreign genetic material (DNA or RNA). ([http://en.wikipedia.org/wiki/Transformation_%28genetics%29 Wikipedia])
Competent cells are bacteria, which can accept extra-chromosomal DNA or plasmids ([http://en.wikipedia.org/wiki/Competent_cells Wikipedia]).
For each plasmid:
- Take competent cells from –80 °C freezer.
- Leave on ice for 10 minutes.
- Add 2-5 μL DNA to competent cell. Remember to label.
- Mix with pipette.
- Sit on ice for 30 minutes.
- Get a beaker of exactly 42 °C water from hot water tap.
- Submerge tubes in hot water for exactly 90 seconds.
- Sit on ice for 5 minutes.
- Add 1 mL of LB.
- Incubate for 1 hour.
- Spread 1 mL on a Petri plate labeled with the appropriate antibiotics. (Dip a glass rod in ethanol and flame it to sterilize, then spread the cells on the plate.)
- Leave the plate with lid half open in the incubator to dry. Rotate the plate 180° every 10 minutes to ensure even spread of bacteria.
- Once dry, turn the plates upside down and incubate overnight.
Jump to [http://igem.skule.ca/lab/protocols/transformation.htm BlueGenes]
