Team:USTC CHINA/Notebook/Protocols/Expression of proteins in Escherichia coli
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2.Add all the 50ml LB medium to an 1L LB medium which has been added 100mg ampicillin and culture the bacterial in a 16℃ incubator shaker until the OD is between 1.0 and 1.2.</br> | 2.Add all the 50ml LB medium to an 1L LB medium which has been added 100mg ampicillin and culture the bacterial in a 16℃ incubator shaker until the OD is between 1.0 and 1.2.</br> | ||
3.Add 3300mM IPTG into the medium and culture the bacterial for 20 to 24 hours.</br> | 3.Add 3300mM IPTG into the medium and culture the bacterial for 20 to 24 hours.</br> | ||
- | 4.Bacteria Collection:Dispense all the bacterial suspension into several 500mL centrifuge bottles and ensure that the error does not exceed 0.1g.Centrifuge under these conditions:The speed is 6000rpm | + | 4.Bacteria Collection:Dispense all the bacterial suspension into several 500mL centrifuge bottles and ensure that the error does not exceed 0.1g.Centrifuge under these conditions:The speed is 6000rpm,the time is 10 minutes and the temperature is 4℃ and discard the supernatant.</br> |
5.Resuspend The bacteria in the centrifuge bottles with 40mL binding buffer and transfer the bacterial suspension to an 80ml beaker for ultrasonic disruption under these conditions:The power is 40%,the total working time is 20min and the on time is 2s while the off time is 4s.</br> | 5.Resuspend The bacteria in the centrifuge bottles with 40mL binding buffer and transfer the bacterial suspension to an 80ml beaker for ultrasonic disruption under these conditions:The power is 40%,the total working time is 20min and the on time is 2s while the off time is 4s.</br> | ||
6.Dispense the bacterial suspension into two 500mL centrifuge bottles and ensure that the error does not exceed 0.1g.Centrifuge under these conditions:The speed is 13000rpm/min,the time is 30 minutes and the temperature is 4℃.</br> | 6.Dispense the bacterial suspension into two 500mL centrifuge bottles and ensure that the error does not exceed 0.1g.Centrifuge under these conditions:The speed is 13000rpm/min,the time is 30 minutes and the temperature is 4℃.</br> |
Revision as of 14:59, 25 September 2013