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<li>Device 3 (number 5) showed a low GFP expression, so it was tried to re-preform the tranformation. Negative control of the interlab (number 1) was not performed last time due to lack of LBC plates so was also performed. </li> | <li>Device 3 (number 5) showed a low GFP expression, so it was tried to re-preform the tranformation. Negative control of the interlab (number 1) was not performed last time due to lack of LBC plates so was also performed. </li> | ||
− | <li><a href = "https://2018.igem.org/Team:Utrecht/Protocol#Transformation"> | + | <li><a href = "https://2018.igem.org/Team:Utrecht/Protocol#Transformation">Protocol Transformation</a></li> |
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Revision as of 21:20, 6 October 2018
PLACEHOLDER
Receptor Assay
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August
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September
BRET Assay
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Methylation Assay
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Morning
Performed Calibration 1, 2, and 3 according to protocol. All measurements were performed using the plate reader of Seino.
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | |
---|---|---|---|---|---|---|---|---|---|---|---|---|
A | L | dd | ||||||||||
B | L | dd | ||||||||||
C | L | dd | ||||||||||
D | L | dd | ||||||||||
E | MS | dd | dd | dd | dd | dd | dd | dd | dd | dd | dd | dd |
F | MS | dd | dd | dd | dd | dd | dd | dd | dd | dd | dd | dd |
G | MS | dd | dd | dd | dd | dd | dd | dd | dd | dd | dd | dd |
H | MS | dd | dd | dd | dd | dd | dd | dd | dd | dd | dd | dd |
Calibration 1
- L= 100 uL Ludox CL-X (stored at 4C)
- dd= 100 uL ddH20
- Measurement: Abs600, turn off pathlength correction
Calibration 2
- MS= 200 ul Microsphere Stock Solution
- dd= 100 uL ddH20
- green= serial dilution was performed with a micropipet from E1,F1,G1,H1 - E11,F11,G11,H11 by a volume of 100uL. Before every transfer solution was pipetted up and down 3x, after every transfer tips were discharged.
- Measurement: Abs600, re-mix befor putting in plate reader and prevent bubbles, path length correction off
Calibration 3
- 1xFC= 200 mL 1xFC (100uL 10x fluorescein + 900ul 1x PBS pH 7.4, tube was covered with foil
- P= 100 uL 1x PBS pH 7.4
- green= serial dilution was performed with a micropipet from A1,B1,C1,D1 - A11,B11,C11,D11 by a volume of 100uL. Before every transfer solution was pipetted up and down 3x, after every transfer tips were discharged.
- Measurement: FL, 530nm/30nm bandpass, 25-30nm with recommened excitation of 485nm, emission 520-530nm of the filter. Path length correction was turned off
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | |
---|---|---|---|---|---|---|---|---|---|---|---|---|
A | 1xFC | P | P | P | P | P | P | P | P | P | P | P |
B | 1xFC | P | P | P | P | P | P | P | P | P | P | P |
C | 1xFC | P | P | P | P | P | P | P | P | P | P | P |
D | 1xFC | P | P | P | P | P | P | P | P | P | P | P |
E | ||||||||||||
F | ||||||||||||
G | ||||||||||||
H |
Afternoon
LBC plates were made according to the protocol used on the wall
- 250ml LB 2x added to melted 250 ml WA 2x using a microwave
- 0.5ml was added to final solution
- plates were dried in 37C incubator
Transformation device 3 + negative control interlab study
- Device 3 (number 5) showed a low GFP expression, so it was tried to re-preform the tranformation. Negative control of the interlab (number 1) was not performed last time due to lack of LBC plates so was also performed.
- Protocol Transformation
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