Team:IISER-Mohali-INDIA/Contribution

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.