IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/07/30: Difference between revisions
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| 14μL of SDW | | 14μL of SDW | ||
| 14μL of SDW | |||
|- | |||
| 2μL of 10X Fast Digest Buffer | | 2μL of 10X Fast Digest Buffer | ||
| 2μL of DNA | | 2μL of Buffer 3 (Biolabs) | ||
|- | |||
| 2μL of DNA of biobricks | |||
| 2μL of DNA of PNZ8901 | |||
|- | |||
| 1μL of EcoRI | | 1μL of EcoRI | ||
| 1μL of PstI | |||
|- | |||
| 1μL of SpeI | | 1μL of SpeI | ||
| 1μL of SalI | | 1μL of SalI | ||
|} | |} |
Revision as of 08:26, 31 July 2008
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Wet Work
Medium A
For this medium, the curb stop to increase logarithmically at 100min.
For this medium, the curb stop to increase logarithmically at 90min.
- Prepare 7 tubes for medium A and 7 tubes for medium A + tryptophan : add 0.45mL of prewarmed medium B and 0.05mL of culture and incubate 90min at 37°C by shaking. - Transform B.S. by adding DNA and incubate 30min at 37°C We have one transformation with ppL82 (add 7.5μL of DNA), one with PNZ8901 (add 10μL of DNA) and one negative control without DNA. - Plate 500μL on Cm plate (35μg/mL)
- Plamsid miniprep (for I746001, I746101 and PNZ8901 from big flasks - Digest
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