Koeris/Notebook/2007-1-8: Difference between revisions
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=Chemically competent bacterial cells= | =Chemically competent bacterial cells= | ||
All cultures are grown at 37 deg C and 300 rpm shaking. The OD<sub>600</sub> of the culture should be between 0.4 - 0.6. The culture | All cultures are grown at 37 deg C and 300 rpm shaking. The OD<sub>600</sub> of the culture should be between 0.4 - 0.6. The culture is then centrifuged down at 3000rpm for 5min and the pellet resuspended in 1/10 volume of TSS. All steps are to be performed on ice. The tubes for the aliquots are also to be pre-chilled. | ||
==Reagents needed== | ==Reagents needed== | ||
The following reagents are needed for the protocol. | The following reagents are needed for the protocol. | ||
Revision as of 07:44, 17 January 2007
Chemically competent bacterial cells
All cultures are grown at 37 deg C and 300 rpm shaking. The OD600 of the culture should be between 0.4 - 0.6. The culture is then centrifuged down at 3000rpm for 5min and the pellet resuspended in 1/10 volume of TSS. All steps are to be performed on ice. The tubes for the aliquots are also to be pre-chilled.
Reagents needed
The following reagents are needed for the protocol.
TSS (Transformation and storage solution)
For 50ml use the following ratios:
- LB: 30ml
- PEG: 5.0g
- DMSO: 2.5ml
- MgCl2[1M]: 0.25ml
- MgSO4[1M]: 0.5ml
- 100% glycerol: 7.5ml
Store all competent cells at -80 deg C for future use.
