Meeting
attendants:
Time:
green light receptor
already done:
To-do:
blue light receptor
already done:
To-do:
red light receptor
already done:
To-do:
Lysis cassette
already done:
To-do:
Precipitator
already done:
To-do:
green light receptor
repeating quickchange PCR
Investigators:Julia
repeating PCR with fresh DNA template.
digest with DpnI,added directly to the PCR tube.
After NEB the DPNI has also a good activity in Phusion HF buffer.
blue light receptor
Ligation
Investigators: Sophie
Date: 25.08.11
continue from experiment: Digestion;
Investigator: Sandra
Project Name: Blue light receptor
all samples stored in Blue light box
Transformation
Investigators: 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
red light receptor
NAME OF YOUR EXPERIMENT
Investigators:NAME
Lysis cassette
NAME OF YOUR EXPERIMENT
Investigators:NAME
Precipitator
NAME OF YOUR EXPERIMENT
Investigators: NAME
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