Team:Freiburg/Notebook/26 August
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===NAME OF YOUR EXPERIMENT=== | ===NAME OF YOUR EXPERIMENT=== | ||
+ | Transformation | ||
+ | Name: Sophie | ||
+ | Date: 26.08.11 | ||
+ | Continue from Date: 26.08.11 Name: Sophie | ||
+ | Experiment Ligation | ||
+ | Project Name: Blue light receptor | ||
- | + | Procedure | |
+ | 1. take cells from -80°C freezer and put them on ice! (every eppi contains about 400 μl cells) | ||
+ | 2. thaw cells on ice 20 minutes | ||
+ | 3. pipette 50 μl cells and 2 μl DNA into eppi still on ice! | ||
+ | 4. Incubate for 30 minutes on ice | ||
+ | 5. Heat at 42°C for 60 sec | ||
+ | 6. Incubate on ice for 5 minutes | ||
+ | 7. Add 200 μl LB Broth | ||
+ | 8. Incubate for 2 hours at 37°C (cells with lysis cassette at 30°C!!) | ||
+ | 9. Plate 50 μl and 200μl on two different LB/Agar plates with appropriate antibiotic resistance | ||
+ | Documentation: | ||
+ | Why are you doing this experiment? Name of the sample? Where are they stored? Name the vector with inserts, antibiotika resistance etc. | ||
+ | Why? The last cloning was done with a NOT-PCR-Product which might make problems because it does not have enough bases after the restriciton sites. | ||
+ | |||
+ | Name of the samples: Not 1:1, Not 1:3, nOt 1:1 nOt 1:3, noT 1:1, noT 1: 3 | ||
+ | |||
+ | stored in incubator on Amp plates | ||
+ | |||
+ | vector: psb1A3 | ||
+ | inserts: Not/ nOt/ noT + LOV-3A-PCR | ||
+ | |||
+ | |||
+ | |||
+ | '''Investigators:NAME''' | ||
==<span style="color:red;">red light receptor</span>== | ==<span style="color:red;">red light receptor</span>== |
Revision as of 12:44, 26 August 2011