Team:IISER-Mohali-INDIA/Contribution

gEco
Contribution






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 exact same protocol around the world to produce common, comparable units for measuring GFP with different plate readers. For more information on the protocol, refer iGEM's official Interlab Study webpage. Below is the data that we collected for the Interlab Study:


OD600 Reference Point Plate Pattern
This diagram depicts how our samples were organized on our plate:
  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
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

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

Curves



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


Parts

Composite Part:

Part no. 1

Part name- Plac->RBS->amilCP Blue chromoprotein

Part number- BBa_K2320001

Description:

AmilCP blue chromoprotein has been cloned under IPTG inducible lac promoter and RBS. Induction of E. coli carrying K2320001 with IPTG leads to intracellular production of blue colored amilCP protein. Other teams can clone their gene of interest along with a RBS of their choice downstream of this IPTG inducible reporter. Induction with IPTG will give a real-time crude estimation of expression of their protein of interest. In addition, this construct will give an added advantage to tune the expression of their protein of interest by varying amount of IPTG or glucose.







Part no. 2

Part name- Plac->RBS->amilGFP BBa_K592010 (yellow)

Part number- BBa_K2320002

Description:

AmilGFP yellow chromoprotein has been cloned under IPTG inducible lac promoter and RBS. Induction of E. coli carrying K2320002 with IPTG leads to intracellular production of yellow colored amilGFP protein. Other teams can clone their gene of interest along with a RBS of their choice downstream of this IPTG inducible reporter. Induction with IPTG will give a real-time crude estimation of expression of their protein of interest. In addition, this construct will give an added advantage to tune the expression of their protein of interest by varying amount of IPTG or glucose.






Part no. 3

Part name- Plac->RBS->amilCP Blue chromoprotein

Part number- BBa_K2320001

Description:

AmilCP blue chromoprotein has been cloned under IPTG inducible lac promoter and RBS. Induction of E. coli carrying K2320001 with IPTG leads to intracellular production of blue colored amilCP protein. Other teams can clone their gene of interest along with a RBS of their choice downstream of this IPTG inducible reporter. Induction with IPTG will give a real-time crude estimation of expression of their protein of interest. In addition, this construct will give an added advantage to tune the expression of their protein of interest by varying amount of IPTG or glucose.







Part no. 4

Part name- Plac->RBS->cjBlue green chromoprotein

Part number- BBa_K2320004

Description:

cjBlue green chromoprotein has been cloned under IPTG inducible lac promoter and RBS. Induction of E. coli carrying K2320004 with IPTG leads to intracellular production of greenish-blue colored chromoprotein. Other teams can clone their gene of interest along with a RBS of their choice downstream of this IPTG inducible reporter. Induction with IPTG will give a real-time crude estimation of expression of their protein of interest. In addition, this construct will give an added advantage to tune the expression of their protein of interest by varying amount of IPTG or glucose.






Part no. 5

Part name- Plac->RBS->eforRed chromoprotein

Part number- BBa_K2320005

Description:

eforRed chromoprotein has been cloned under IPTG inducible lac promoter and RBS. Induction of E. coli carrying K2320005 with IPTG leads to intracellular production of eforRed chromoprotein. Other teams can clone their gene of interest along with a RBS of their choice downstream of this IPTG inducible reporter. Induction with IPTG will give a real-time crude estimation of expression of their protein of interest. In addition, this construct will give an added advantage to tune the expression of their protein of interest by varying amount of IPTG or glucose.