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Revision as of 06:45, 20 October 2009

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Transformation Protocol

From iGEM Plates

1. Add 15µl TE (Tris-EDTA, pH 8.0) to well containing part and pipette up and down to resuspend
2. Take 1µl of the above solution and mix with partly thawed competent cells on ice
3. Leave on ice for 30 min.
4. Heat Shock for at 42C for 45 seconds
5. Return eppendorf containing cells to ice and leave for 5 min.
6. Add 500µl LB and culture at 37C for 30 min.
7. Spread on plate with appropriate antibiotic resistance
8. Culture plate at 37C overnight

After ligation or from miniprep

1. Take 1µl(if from miniprep)/5µl(if from ligation) of DNA solution and mix with partly thawed competent cells on ice
2. Leave on ice for 30 min.
3. Heat Shock for at 42C for 45 seconds
4. Return eppendorf containing cells to ice and leave for 5 min.
5. Add 500µl LB and culture at 37C for 30 min.
6. Spread on plate with appropriate antibiotic resistance
7. Culture plate at 37C overnight

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