green light receptor
Digest of Qickchange PCR product with DpnI
Investigators:Julia
in order to get rid of old plasmids a digest with DpnI was performed.
Transformation
Competent cells (50µl) were transformed with 10µl of the PCR- Product after iGEM Protocol.
CcaS is in the iGEM vector pSB1K3
CcaR in pSB1C3.
blue light receptor
Digestion of PCR product
Investigators:Sandra
Digest of PCR product S35 (Lov-tap), because PCR did not work out. We got a band of 1000bp, but we expected 3000bp.
Digest:
- 1 μl NEB4 buffer
- 1 μl BSA
- 1 μl NcoI
- 2 μl H2O
- 5 μl S35
Incubation for 30 min 37°C.
Afterwards the digested product was loaded onto gel.
Comments: No results. We will try a gradient PCR.
Gradient PCR for Lov-tap S35
Investigators: Sophie
Comments: Because PCR for S35 did not work, we are doing a gradient PCR.
PCR as usual. Only temperature were adjusted, to determine the optimum of the annealing temperature.
Miniprep of Not-Gate
Investigators: Sophie
Picked colonies from plate and then performed a miniprep.
DNA-concentration measured with nanodrop:
| sample
| DNA-concentration (ng/μl)
|
| M45a
| 248.9
|
| M45b
| 256.8
|
| M45c
| 291.3
|
| M45d
| 295.8
|
Testdigest of Miniprep
Investigators: Sophie
Testdigest of M45a-M45d.
- 4 μl H20
- 1 μl NEB4
- 1 μl BSA
- 0.5 μl EcoRI
- 0.5 μl PStI
- 3 μl DNA (M45a-M45d)
Incubation for 1 hour. Gel was loaded afterwards.
red light receptor
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Lysis cassette
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Precipitator
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