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<h1>Calibration Protocol</h1> | <h1>Calibration Protocol</h1> | ||
<p>Three calibration measurement were done by the calibration protocol, including an OD reference point at 600nm, one particle standard curve and one fluorescent standard curve.</p> | <p>Three calibration measurement were done by the calibration protocol, including an OD reference point at 600nm, one particle standard curve and one fluorescent standard curve.</p> | ||
− | < | + | <h2>★ OD600 reference point ★ </h2> |
− | + | <br> <br> | |
− | + | <table border="1"> | |
− | <br> <br> | + | <thead> |
− | <table border="1"> | + | <tr> |
− | + | <th></th> | |
− | + | <th>LUDOX CL-X</th> | |
− | + | <th>H2O</th> | |
− | + | </tr> | |
− | + | </thead> | |
− | + | <tbody> | |
− | </thead> | + | <tr> |
− | + | <th>Replicate 1</th> | |
− | + | <th class="bluetable">0.066</th> | |
− | + | <th class="bluetable">0.0285</th> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <th>Replicate 2</th> | |
− | + | <th>0.059</th> | |
− | + | <th>0.029</th> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <th>Replicate 3</th> | |
− | + | <th>0.068</th> | |
− | + | <th>0.026</th> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <th>Replicate 4</th> | |
− | + | <th>0.064</th> | |
− | + | <th>0.028</th> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <th>Arith. Mean</th> | |
− | + | <th>0.064</th> | |
− | + | <th>0.028</th> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <th>Corrected Abs600</th> | |
− | + | <th> 0.036</th> | |
− | + | <th></th> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <th>Reference OD600</th> | |
− | + | <th>OD600</th> | |
− | + | <th></th> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <th>OD600/Abs600</th> | |
− | + | <th>1.728</th> | |
− | + | <th></th> | |
− | + | </tr> | |
− | + | </tbody> | |
− | + | </table> | |
− | + | <img src="https://static.igem.org/mediawiki/2018/4/42/T--ZJUT-China--Particle_Standard_Curve.png" class="col-2" alt="Particle_Standard_Curve.png"> | |
− | </table> | + | <img src="https://static.igem.org/mediawiki/2018/f/fc/T--ZJUT-China--Particle_Standard_Curve_%28log_scale%29e.png" class="col-2"alt="Particle_Standard_Curve_(log_scale)e"> |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
</div> | </div> | ||
<div class="column judges-will-not-evaluate middle" > | <div class="column judges-will-not-evaluate middle" > |
Revision as of 18:43, 24 July 2018
INTERLAB
Our team participated in the Fifth International InterLaboratory Measurement Study this year. The interlab study aims to solve the problem of reliability and repeatability in synthetic biology study. This year we were asked to measure green fluorescent protein, which is used as a measurement marker. We followed the experiment protocol strictly to make sure our data is valid. After two weeks lab work, we got the data we need after several attempts.
Calibration Protocol
Three calibration measurement were done by the calibration protocol, including an OD reference point at 600nm, one particle standard curve and one fluorescent standard curve.
★ OD600 reference point ★
LUDOX CL-X | H2O | |
---|---|---|
Replicate 1 | 0.066 | 0.0285 |
Replicate 2 | 0.059 | 0.029 |
Replicate 3 | 0.068 | 0.026 |
Replicate 4 | 0.064 | 0.028 |
Arith. Mean | 0.064 | 0.028 |
Corrected Abs600 | 0.036 | |
Reference OD600 | OD600 | |
OD600/Abs600 | 1.728 |
★ Fluorescence Raw Readings:★
Hour 0: | Neg. Control | Pos. Control | Device 1 | Device 2 | Device 3 | Device 4 | Device 5 | Device 6 | LB + Chlor (blank) |
---|---|---|---|---|---|---|---|---|---|
Colony 1, Replicate 1 | 98.891 | 514.509 | 170.45 | 453.211 | 107.527 | 377.998 | 110.335 | 121.276 | 100.56 |
Colony 1, Replicate 2 | 87.951 | 520.569 | 155.638 | 443.722 | 103.159 | 353.38 | 99.569 | 117.851 | 97.983 |
Colony 1, Replicate 3 | 89.665 | 500.61 | 151.21 | 474.651 | 99.334 | 358.843 | 97.329 | 115.674 | 97.686 |
Colony 1, Replicate 4 | 88.651 | 553.489 | 158.106 | 471.047 | 107.222 | 391.116 | 102.558 | 132.724 | 104.901 |
Colony 2, Replicate 1 | 94.201 | 799.161 | 176.469 | 396.183 | 105.65 | 452.456 | 105.862 | 129.379 | 100.254 |
Colony 2, Replicate 2 | 97.455 | 750.414 | 168.71 | 425.555 | 106.286 | 462.598 | 109.024 | 125.311 | 101.914 |
Colony 2, Replicate 3 | 95.836 | 795.354 | 174.674 | 405.952 | 103.704 | 444.673 | 106.365 | 128.606 | 105.761 |
Colony 2, Replicate 4 | 100.733 | 804.913 | 203.631 | 370.775 | 117.227 | 506.476 | 113.511 | 125.021 | 105.18 |
Hour 6: | Neg. Control | Pos. Control | Device 1 | Device 2 | Device 3 | Device 4 | Device 5 | Device 6 | LB + Chlor (blank) |
---|---|---|---|---|---|---|---|---|---|
Colony 1, Replicate 1 | 142.426 | 1579.487 | 407.651 | 2268.46 | 167.703 | 1014.837 | 144.06 | 365.966 | 130.779 |
Colony 1, Replicate 2 | 143.238 | 1593.226 | 396.943 | 2328.22 | 174.862 | 1003.434 | 141.231 | 339.499 | 126.638 |
Colony 1, Replicate 3 | 151.515 | 1699.317 | 396.669 | 2175.501 | 169.71 | 977.6 | 143.762 | 347.711 | 127.911 |
Colony 1, Replicate 4 | 155.799 | 1783.066 | 406.205 | 2240.128 | 171.295 | 1044.087 | 154.138 | 340.887 | 127.237 |
Colony 2, Replicate 1 | 150.46 | 2062.959 | 354.432 | 2090.23 | 165.189 | 1268.176 | 150.674 | 320.936 | 120.878 |
Colony 2, Replicate 2 | 145.579 | 2023.17 | 366.367 | 2169.758 | 167.611 | 1203.787 | 144.458 | 322.957 | 127.103 |
Colony 2, Replicate 3 | 146.553 | 2101.747 | 353.019 | 1926.808 | 183.724 | 1172.553 | 158.135 | 332.951 | 131.201 |
Colony 2, Replicate 4 | 157.503 | 2071.459 | 316.402 | 1750.215 | 179.275 | 1253.449 | 155.625 | 327.385 | 137.828 |
★ Abs600 Raw Readings:★
Hour 0: | Neg. Control | Pos. Control | Device 1 | Device 2 | Device 3 | Device 4 | Device 5 | Device 6 | LB + Chlor (blank) |
---|---|---|---|---|---|---|---|---|---|
Colony 1, Replicate 1 | 0.1306 | 0.0884 | 0.0767 | 0.1068 | 0.1181 | 0.0753 | 0.1067 | 0.0866 | 0.0486 |
Colony 1, Replicate 2 | 0.1272 | 0.0906 | 0.0737 | 0.1075 | 0.1184 | 0.0754 | 0.108 | 0.0896 | 0.0497 |
Colony 1, Replicate 3 | 0.1246 | 0.09 | 0.0747 | 0.1119 | 0.1214 | 0.0731 | 0.1083 | 0.0879 | 0.0464 |
Colony 1, Replicate 4 | 0.1225 | 0.0872 | 0.0743 | 0.1109 | 0.1247 | 0.0746 | 0.1079 | 0.0903 | 0.0476 |
Colony 2, Replicate 1 | 0.1301 | 0.0907 | 0.0701 | 0.1109 | 0.1132 | 0.0813 | 0.1103 | 0.0909 | 0.0468 |
Colony 2, Replicate 2 | 0.1327 | 0.0919 | 0.0685 | 0.1125 | 0.1145 | 0.0822 | 0.1081 | 0.0896 | 0.0472 |
Colony 2, Replicate 3 | 0.1294 | 0.0952 | 0.0693 | 0.1151 | 0.1139 | 0.0809 | 0.1084 | 0.0898 | 0.05 |
Colony 2, Replicate 4 | 0.1202 | 0.0941 | 0.0692 | 0.1105 | 0.1134 | 0.0805 | 0.1092 | 0.089 | 0.0472 |
Hour 6: | Neg. Control | Pos. Control | Device 1 | Device 2 | Device 3 | Device 4 | Device 5 | Device 6 | LB + Chlor (blank) |
---|---|---|---|---|---|---|---|---|---|
Colony 1, Replicate 1 | 0.9386 | 0.7564 | 0.1978 | 0.911 | 0.7963 | 0.2566 | 0.8131 | 0.6363 | 0.0444 |
Colony 1, Replicate 2 | 0.9413 | 0.7528 | 0.1974 | 0.9012 | 0.8211 | 0.2547 | 0.7951 | 0.5996 | 0.0438 |
Colony 1, Replicate 3 | 0.9745 | 0.7748 | 0.1951 | 0.8642 | 0.81 | 0.2536 | 0.7994 | 0.6044 | 0.0458 |
Colony 1, Replicate 4 | 0.9932 | 0.7975 | 0.2009 | 0.8969 | 0.8021 | 0.2622 | 0.8266 | 0.6092 | 0.0438 |
Colony 2, Replicate 1 | 0.8246 | 0.78 | 0.1031 | 0.8625 | 0.8193 | 0.2834 | 0.7633 | 0.5251 | 0.0457 |
Colony 2, Replicate 2 | 0.8327 | 0.7665 | 0.1031 | 0.9018 | 0.8211 | 0.2781 | 0.7394 | 0.5374 | 0.0462 |
Colony 2, Replicate 3 | 0.8062 | 0.7927 | 0.1005 | 0.829 | 0.8082 | 0.276 | 0.7689 | 0.5668 | 0.0458 |
Colony 2, Replicate 4 | 0.8262 | 0.7805 | 0.1005 | 0.8787 | 0.8156 | 0.2928 | 0.7604 | 0.5332 | 0.0477 |
★ Assumed plate well pattern:★
A1 | A2 | A3 | A4 | A5 | A6 | A7 | A8 | A9 |
---|---|---|---|---|---|---|---|---|
B1 | B2 | B3 | B4 | B5 | B6 | B7 | B8 | B9 |
C1 | C2 | C3 | C4 | C5 | C6 | C7 | C8 | C9 |
D1 | D2 | D3 | D4 | D5 | D6 | D7 | D8 | D9 |
E1 | E2 | E3 | E4 | E5 | E6 | E7 | E8 | E9 |
F1 | F2 | F3 | F4 | F5 | F6 | F7 | F8 | F9 |
G1 | G2 | G3 | G4 | G5 | G6 | G7 | G8 | G9 |
H1 | H2 | H3 | H4 | H5 | H6 | H7 | H8 | H9 |