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:(C-colony, R-replicate) |
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 |