Morning

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

Table 1. content 96 wells plate cal.1 and cal.2

	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

Table 2. content 96 wells plate cal.3

	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

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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