Difference between revisions of "Team:Aix-Marseille/Experiments/Protocols"
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Revision as of 12:08, 9 June 2017
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Contents
Protocols
Preparation of competent Bacterial cells
1- Preparation of the Bacterial culture
- Recuperate the overnight bacterial culture
- Determine the OD600 in a 1 ml Tank Spectro, dilute a 100µl of culture in 900µl of distilled LB
- In 1L Erlenmeyer add 200 ml of LB and an appropriate volume of culture to have an OD600 = 0.1
- Put the Erlenmeyer in an incubator for about an hour at 37˚C.
- test the OD600 for the new culture
- the OD600 should be between 0.4 and 0.6
All the handling done in a 15cm radius of an open flame for optimal sterility
2- Preparation of Buffer Tbf1 and Tbf2
| Tbf1 buffer | Total volume 80mL |
| KAc 1M | 2.4 mL |
| MnCl2 0.5M | 8 mL |
| KCl 1M | 8 mL |
| CaCl2 0.1M | 8 mL |
| GlY 80% | 15mL |
| H2O | 38.6 mL |
