Morning

Performed Calibration 1, 2, and 3 according to protocol. All measurements were performed using the plate reader of Seino.

Table 2: Receptor EC50 values

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 07/08/18 (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 used: competent cells (DH5-a) was put on ice for 20 min 50ul of competent cells were transferred to inoculation tube, one for each separate DNA sample 2ul DNA (number 5 or 1) was added 30 min on ice 42 C heatshock for 45 sec coldshock on ice 5 min 500uL SOB was added to each of the tubes 1h incubated in 37C shaker plated on LBC plates and incubated overnight at 37 C 100ul directly (with resuspending) centrifugation at full speed for 30 sec 300ul was removed pellet was resuspended in remaining 100ul 100ul was plated onto LBC plate

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