green light receptor
Ligation of Promotor PcpcG in PSB1C3
Investigators:Julia
1. Digestion of PCR Product from yesterday[1]
38µl PCR product
5µl BSA (10x)
5µl NEB buffer 4
1µl EcoRI
1µlPstI
Incubation for 3 hours, afterwards the sample was cleaned with a PCR purification kit from Qiagen.
2.Ligation
11µl H2O
4µl PcpcG
2µl pSB1C3
2µl T4 Ligase buffer
1µl T4 Ligase
Incubation for 3 hours at room temperature.
heat inactivation at 80 degrees for 20 min.
3. transformation
blue light receptor
Transformation
...with ♥-A3-Not, ♥-A3-nOt and ♥-A3-noT </br>
Investigators: Sophie
red light receptor
Investigators:Julia
Lysis cassette
NAME OF YOUR EXPERIMENT
Investigators:NAME
Precipitator
Cloning
Investigators: Sophie
Project name: inducible promoter for pbd
again cloning of IPTG-inducible Promoter, new psb1C3-PCR product and ε5 (GFP-pbd) but this time I use Antarctic Phosphatase in the last hour of the digestion of the vector to avoid re-ligation. For 50μL reaction volume use 5,6μL Antarctic Phosphatase buffer and 1μL Antarctic Phosphatase.
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