Team:Todai-Tokyo/Protocols/Transformation
From 2009.igem.org
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Transformation Protocol
From iGEM Plates
- Add 15µl TE (Tris-EDTA, pH 8.0) to well containing part and pipette up and down to resuspend
- Take 1µl of the above solution and mix with partly thawed competent cells on ice
- Leave on ice for 30 min.
- Heat Shock for at 42ºC for 45 seconds
- Return eppendorf containing cells to ice and leave for 5 min.
- Add 500µl LB and culture at 37C for 30 min.
- Spread on plate with appropriate antibiotic resistance
- Culture plate at 37ºC overnight
After ligation or from miniprep
- Take 1µl (if from miniprep)/5µl (if from ligation) of DNA solution and mix with partly thawed competent cells on ice
- Leave on ice for 30 min.
- Heat Shock for at 42ºC for 45 seconds
- Return eppendorf containing cells to ice and leave for 5 min.
- Add 500µl LB and culture at 37ºC for 30 min.
- Spread on plate with appropriate antibiotic resistance
- Culture plate at 37ºC overnight