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Revision as of 03:52, 10 October 2017


Interlab

iGEM's interlab study aims to establish a GFP measurement protocol based on engineering principles that anyone with a plate reader can use in their lab. This means teams have to use the same exact protocol around the world to produce common, comparable units for measuring GFP with different plate readers. For more information on the protocol, check out iGEM's official Interlab Study webpage. Below is the data that we collected for the Interlab Study:


OD600 Reference Point

  LUDOX-HS40 H2O
Replicate 1 0.0932 0.0973
Replicate 2 0.0927 0.0777
Replicate 3 0.0961 0.0796
Replicate 4 0.0937 0.0803
Arith. Mean 0.093925 0.083725
Corrected Abs600 0.0102
Reference OD600 0.0425
OD600/Abs600 4.166666667

Plate Pattern

This diagram depicts how our samples were organized on our plate:

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

Fluorescence Standard Curves


uM Fluorescein

uM Fluorescein 50.00 25 12.5 6.25 3.125 1.5625 0.78125 0.390625 0.1953125 0.09765625 0.048828125 0
Replicate 1 60105 43268 29004 17671 9656 4867 2466 1328 658 324 160 6
Replicate 2 64328 45105 30375 18411 9892 4742 2569 1361 698 339 172 6
Replicate 3 64879 46230 30542 18336 9472 4772 2540 1353 703 349 179 5
Replicate 4 64574 46956 30906 18065 9491 5071 2587 1374 709 359 190 6
Arith. Mean 63471.5 45389.75 30206.75 18120.75 9627.75 4863 2540.5 1354 692 342.75 175.25 5.75
Arith. Std.Dev. 2255.620905 1606.452069 831.9172134 334.6444631 194.5805317 148.550777 53.30728531 19.37352145 23.10844002 14.93039406 12.57974563 0.5

uM Fluorescein/a.u.


uM Fluorescein/a.u. 50.00 25 12.5 6.25 3.125 1.5625 0.78125 0.390625 0.1953125 0.09765625 0.048828125
Mean um Fluorescein/a.u. 0.000787755 0.000550785 0.000413815 0.000344908 0.000324583 0.000321304 0.000307518 0.000288497 0.000282243 0.00028492 0.00027862
Mean of med-high levels: 0.000391079

Abs600 Raw Readings

These tables summarize our finalized sets of Absorbance reading data over an incubation period of 6 hours with samples measured every 2 hours for a total of 4 time points:

Hour 0

Hour 0: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6 LB + Chlor (blank)
C1, R1 0.0935 0.0951 0.0942 0.0917 0.0918 0.0925 0.0919 0.0933 0.0902
C1, R2 0.0937 0.0937 0.0973 0.0941 0.0937 0.096 0.0937 0.0945 0.0886
C1, R3 0.0946 0.0939 0.1013 0.0957 0.0931 0.1026 0.1074 0.0945 0.0904
C1, R4 0.0982 0.0943 0.0996 0.0942 0.0926 0.0939 0.094 0.0953 0.0899
C2, R1 0.0932 0.0944 0.0956 0.0933 0.0927 0.0919 0.0934 .0973 0.0894
C2, R2 0.0928 0.0949 0.0975 0.0967 0.0976 0.0949 0.0962 0.0957 0.0894
C2, R3 0.0925 0.094 0.0983 0.095 0.0939 0.0941 0.0946 0.0955 0.0916
C2, R4 0.0918 0.0965 0.0958 0.0942 0.0939 0.0923 0.093 0.0932 0.0887

Hour 2

Hour 2: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6 LB + Chlor (blank)
C1, R1 0.1289 0.1165 0.0932 0.1002 0.1056 0.0952 0.1129 0.1145 0.0892
C1, R2 0.1294 0.1158 0.0975 0.1031 0.1087 0.0973 0.1136 0.117 0.0922
C1, R3 0.1305 0.1156 0.0962 0.1013 0.108 0.0982 0.1157 0.1174 0.0916
C1, R4 0.1275 0.1153 0.096 0.1019 0.11 0.0964 0.1115 0.1189 0.0845
C2, R1 0.1273 0.1221 0.0957 0.1027 0.1165 0.0956 0.1147 0.1118 0.0844
C2, R2 0.129 0.1192 0.0958 0.1033 0.1198 0.0935 0.1162 0.114 0.0922
C2, R3 0.1246 0.1221 0.0961 0.1021 0.1178 0.096 0.1156 0.1114 0.0921
C2, R4 0.1258 0.1227 0.0969 0.0995 0.121 0.096 0.1139 0.1134 0.0895

Hour 4

Hour 4: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6 LB + Chlor (blank)
C1, R1 0.1906 0.1708 0.0939 0.1434 0.1767 0.1029 0.1698 0.1784 0.0908
C1, R2 0.1923 0.1717 0.0953 0.1479 0.179 0.1045 0.1761 0.1805 0.0904
C1, R3 0.1951 0.1747 0.1072 0.1504 0.1809 0.1049 0.1741 0.184 0.0911
C1, R4 0.1996 0.1797 0.0979 0.1519 0.1833 0.109 0.1788 0.1824 0.0906
C2, R1 0.1889 0.1833 0.0961 0.1369 0.1802 0.0958 0.1703 0.1872 0.0902
C2, R2 0.1903 0.1844 0.0986 0.1368 0.1863 0.0988 0.171 0.1956 0.092
C2, R3 0.189 0.1891 0.0971 0.139 0.1821 0.0974 0.1701 0.1874 0.0958
C2, R4 0.1891 0.1865 0.0953 0.1417 0.1815 0.0976 0.167 0.1845 0.0895

Hour 6

Hour 6: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6 LB + Chlor (blank)
C1, R1 0.2852 0.2515 0.0937 0.2081 0.2876 0.1312 0.2516 0.2616 0.0902
C1, R2 0.2722 0.2545 0.0956 0.2188 0.2918 0.1308 0.2483 0.2663 0.0907
C1, R3 0.2719 0.2535 0.0956 0.2184 0.2929 0.1328 0.2542 0.2714 0.0912
C1, R4 0.2815 0.2591 0.0971 0.2148 0.2959 0.1347 0.2488 0.2649 0.0905
C2, R1 0.2634 0.2673 0.0954 0.1858 0.2583 0.102 0.2342 0.2635 0.0901
C2, R2 0.2651 0.2679 0.097 0.1857 0.2638 0.1042 0.2372 0.2719 0.0926
C2, R3 0.268 0.2681 0.0952 0.1875 0.2624 0.1047 0.2326 0.2628 0.0928
C2, R4 0.2525 0.2702 0.1024 0.1837 0.2582 0.1034 0.2384 0.2648 0.0901

Fluorescence Raw Readings

These tables summarize our finalized sets of Fluorescent reading data over an incubation period of 6 hours:

Hour 0

Hour 0: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6 LB + Chlor (blank)
C1, R1 63 69 130 83 64 76 66 64 64
C1, R2 63 72 129 84 64 75 70 65 64
C1, R3 64 70 131 84 65 76 66 66 67
C1, R4 64 72 132 84 65 77 67 65 65
C2, R1 63 71 124 81 63 74 68 65 66
C2, R2 63 71 122 82 63 74 65 65 68
C2, R3 64 71 123 82 64 75 67 63 67
C2, R4 64 71 123 83 64 75 65 65 66

Hour 2

Hour 2: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6 LB + Chlor (blank)
C1, R1 67 112 135 135 70 97 94 65 65
C1, R2 68 113 139 134 71 98 96 67 67
C1, R3 66 113 142 137 69 97 98 68 66
C1, R4 66 115 136 139 71 95 95 65 70
C2, R1 65 122 133 128 67 86 95 66 70
C2, R2 64 121 136 129 66 84 97 65 65
C2, R3 63 118 133 131 66 86 97 65 66
C2, R4 63 117 136 129 67 86 98 64 65

Hour 4

Hour 4: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6 LB + Chlor (blank)
C1, R1 67 170 153 235 76 118 126 70 65
C1, R2 69 173 150 236 77 116 128 71 66
C1, R3 68 179 156 237 76 117 125 71 66
C1, R4 69 178 155 235 76 121 128 70 67
C2, R1 66 175 145 216 70 91 124 72 67
C2, R2 69 176 143 209 72 88 124 69 65
C2,3 66 176 145 219 70 92 124 70 66
C2, R4 68 177 145 216 73 90 123 72 66

Hour 6

Hour 6: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6 LB + Chlor (blank)
C1, R1 72 256 154 385 85 147 161 77 65
C1, R2 72 263 158 388 88 146 161 76 66
C1, R3 71 266 158 395 87 149 164 77 66
C1, R4 73 272 160 379 84 149 160 73 67
C2, R1 72 259 148 325 78 93 148 74 66
C2, R2 71 258 147 321 77 96 152 75 64
C2, R3 70 259 146 323 79 97 151 76 66
C2, R4 69 259 143 315 79 97 146 75 66