Line 30: | Line 30: | ||
firstMonth = 'A'; | firstMonth = 'A'; | ||
inactiveJuly = []; | inactiveJuly = []; | ||
− | inactiveAugust = [ | + | inactiveAugust = [1,2,3,4,5,6,7,]; |
inactiveSeptember = [15, 16, 19]; | inactiveSeptember = [15, 16, 19]; | ||
firstInactive = inactiveAugust; | firstInactive = inactiveAugust; | ||
Line 259: | Line 259: | ||
<div class = "notebookContainer"> | <div class = "notebookContainer"> | ||
− | <div class="customelementM5B" id = " | + | <div class="customelementM5B" id = "entrydayIA1"> |
− | + | ||
+ | <h3> Morning </h3> | ||
+ | |||
+ | Performed Calibration 1, 2, and 3 according to protocol. All measurements were performed using the plate reader of Seino. | ||
+ | |||
+ | <table class="InterlabTable"> | ||
+ | <caption style = "font-size: 1.5em;background-color: #A4C2F4">Table 2: Receptor EC50 values</caption> | ||
+ | <tr><th>1</th><th>2</th><th>3</th><th>4</th><th>5</th><th>6</th><th>7</th><th>8</th><th>9</th><th>10</th><th>11</th><th>12</th></tr> | ||
+ | <tr><td>A</td><td> L</td><td> dd</td></tr> | ||
+ | <tr><td>B</td><td> L</td><td> dd </td></tr> | ||
+ | <tr><td>C</td><td> L</td><td> dd </td></tr> | ||
+ | <tr><td>D</td><td> L</td><td> dd </td></tr> | ||
+ | <tr><td>E</td><td style="color: green"> MS</td><td style="color: green">dd</td><td style="color: green">dd</td><td style="color: green">dd</td><td style="color: green"> dd</td><td style="color: green"> dd</td><td style="color: green"> dd</td><td style="color: green"> dd</td><td style="color: green"> dd</td><td style="color: green"> dd</td><td style="color: green"> dd</td><td style="color: green"> dd</td></tr> | ||
+ | <tr><td>F</td><td style="color: green"> MS</td><td style="color: green">dd</td><td style="color: green">dd</td><td style="color: green">dd</td><td style="color: green"> dd</td><td style="color: green"> dd</td><td style="color: green"> dd</td><td style="color: green"> dd</td><td style="color: green"> dd</td><td style="color: green">dd</td><td style="color: green">dd</td><td style="color: green">dd</td></tr> | ||
+ | <tr><td>G</td><td style="color: green"> MS</td><td style="color: green">dd</td><td style="color: green">dd</td><td style="color: green">dd</td><td style="color: green">dd</td><td style="color: green">dd</td><td style="color: green">dd</td><td style="color: green">dd</td><td style="color: green">dd</td><td style="color: green">dd</td><td style="color: green">dd</td><td style="color: green">dd</td></tr> | ||
+ | <tr><td>H</td><td style="color: green"> MS</td><td style="color: green">dd</td><td style="color: green">dd</td><td style="color: green">dd</td><td style="color: green">dd</td><td style="color: green">dd</td><td style="color: green">dd</td><td> dd</td><td style="color: green">dd</td><td style="color: green">dd</td><td style="color: green">dd<</td><td style="color: green">dd</td></tr> | ||
+ | </table> | ||
+ | |||
+ | |||
+ | 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 | ||
+ | |||
+ | |||
</div> | </div> | ||
Revision as of 20:38, 6 October 2018
PLACEHOLDER
Interlab
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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 | |
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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 |
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