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Team:Freiburg/Notebook/28 July
From 2011.igem.org
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*3 μl DNA (M45a-M45d)<br/> | *3 μl DNA (M45a-M45d)<br/> | ||
| - | Incubation for 1 hour. Gel was loaded afterwards. | + | Incubation for 1 hour. Gel was loaded afterwards.<br/> |
| - | + | Comments: M45c and M45d look good. | |
[[File:7_29_2011_9_52_26PM.jpg|caption|border]] | [[File:7_29_2011_9_52_26PM.jpg|caption|border]] | ||
Revision as of 12:22, 28 July 2011
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Contents |
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: Not expected 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.
- 54-59°C
- 64-72°C
After the PCR we will load the samples onto a gel to see the results of the gradient PCR.
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.
Comments: M45c and M45d look good.
red light receptor
Miniprep of BBa_I15008 and BBa_I15009
Investigators:Julia
because there was some confusion with the labbeling and no stock, I am repeating transformation and DNA extraction.
Lysis cassette
NAME OF YOUR EXPERIMENT
Investigators:NAME
Precipitator
NAME OF YOUR EXPERIMENT
Investigators: NAME
