Team:ColumbiaNYC/InterLab

Interlab

iGEM's interlab study aims to establish a baseline and robust protocol for fluorescent measurements. Teams that participate in the interlab study will follow a standardized procedure that guides them through the transformation, inoculation and measurement processes. For more information on the protocol, check out iGEM's official Interlab Study webpage. Feel free to also refer to our Lab Notebook, which also lays out what we did for the Interlab Study. Below is the data that we collected for the Interlab Study:

OD600 Reference Point

Several measurements of LUDOX-S40 and distilled water were taken with a spectrophotometer. The resulting data was then analyzed to set an OD600 Reference point and calculate the correction factor, which was applied to the final set of absorbance data in the transformed E. coli DH5 alpha cells
LUDOX-HS40 H2O
Replicate 1 0.097 0.082
Replicate 2 0.092 0.08
Replicate 3 0.09 0.082
Replicate 4 0.091 0.082
Arith. Mean 0.0925 0.0815
Corrected Abs600 0.011
Reference OD600 0.0425
OD600/Abs600 3.863636364

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 35189 28288 20495 16652 12162 8151 8016 8072 8362 7632 5075 8434
Replicate 2 34588 27070 18993 18141 11546 10531 8528 8507 7813 7638 5168 8517
Replicate 3 34302 28580 20980 16855 11988 9524 9391 8676 8107 7997 5116 8691
Replicate 4 34386 27353 21253 16614 11953 10064 8787 8667 7569 7550 5128 8053
Arith. Mean 34616.25 27822.75 20430.25 17065.5 11912.25 9567.5 8680.5 8480.5 7962.75 7704.25 5121.75 8423.75
Arith. Std.Dev. 400.2535655 725.0707437 1008.13735 724.7631797 260.7123255 1030.082359 571.830103 283.1825089 345.2886474 199.2525617 38.28293789 269.3688611

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. 0.001444408 0.000898545 0.000611838 0.000366236 0.000262335 0.000163313 9.00006E-05 4.60616E-05 2.45283E-05 1.26756E-05 9.53348E-06
Mean of med-high levels: 0.000460453

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 0.109 0.104 0.111 0.106 0.096 0.1455 0.107 0.106 0.0865
C1, R2 0.114 0.1185 0.1065 0.107 0.097 0.148 0.109 0.108 0.088
C1, R3 0.109 0.108 0.108 0.108 0.094 0.1465 0.109 0.107 0.086
C1, R4 0.1085 0.106 0.105 0.1065 0.096 0.1485 0.11 0.1095 0.091
C2, R1 0.108 0.1035 0.1105 0.1055 0.1305 0.0985 0.11 0.1145 0.087
C2, R2 0.109 0.105 0.111 0.1085 0.133 0.095 0.3745 0.114 0.086
C2, R3 0.1105 0.105 0.1115 0.1075 0.132 0.1325 0.272 0.4605 0.097
C2, R4 0.1115 0.108 0.112 0.107 0.132 0.095 0.108 0.1155 0.643

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 0.247 0.221 0.1125 0.1955 0.1965 0.2175 0.206 0.238 0.084
C1, R2 0.2315 0.206 0.1125 0.222 0.1925 0.207 0.2125 0.224 0.086
C1, R3 0.227 0.2075 0.1115 0.2145 0.195 0.2195 0.2135 0.2135 0.085
C1, R4 0.227 0.1975 0.1135 0.215 0.1935 0.2155 0.193 0.212 0.084
C2, R1 0.2295 0.204 0.1105 0.191 0.3015 0.179 0.209 0.2775 0.085
C2, R2 0.2325 0.2065 0.111 0.201 0.312 0.1805 0.1965 0.2665 0.085
C2, R3 0.225 0.19 0.1085 0.1905 0.293 0.163 0.174 0.242 0.0835
C2, R4 0.227 0.1915 0.108 0.1995 0.2955 0.1595 0.1965 0.2435 0.086

Hour 4

Hour 4: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6 LB + Chlor (blank)
Colony 1, Replicate 1 0.5255 0.549 0.13 0.54 0.4745 0.6265 0.5025 0.665 0.085
Colony 1, Replicate 2 0.465 0.505 0.124 0.5185 0.4115 0.4235 0.4905 0.569 0.099
Colony 1, Replicate 3 0.5155 0.5 0.126 0.51 0.4425 0.4875 0.501 0.532 0.084
Colony 1, Replicate 4 0.5115 0.5075 0.125 0.5105 0.457 0.4195 0.523 0.535 0.083
Colony 2, Replicate 1 0.5095 0.5055 0.117 0.519 0.626 0.407 0.49 0.696 0.085
Colony 2, Replicate 2 0.4635 0.4935 0.11 0.5005 0.5975 0.454 0.4475 0.562 0.085
Colony 2, Replicate 3 0.5005 0.496 0.1145 0.432 0.6055 0.4525 0.506 0.5465 0.085
Colony 2, Replicate 4 0.506 0.5 0.114 0.488 0.583 0.429 0.562 0.5665 0.084

Hour 6

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 0.681 0.7495 0.198 0.7775 0.781 0.7615 0.729 0.7285 0.087
Colony 1, Replicate 2 0.6595 0.7035 0.168 0.734 0.6505 0.7015 0.7275 0.6735 0.088
Colony 1, Replicate 3 0.6635 0.7295 0.167 0.752 0.659 0.7205 0.419 0.7025 0.085
Colony 1, Replicate 4 0.655 0.724 0.162 0.708 0.727 0.6975 0.6985 0.684 0.085
Colony 2, Replicate 1 0.666 0.957 0.1645 0.6915 0.747 0.719 0.962 0.944 0.085
Colony 2, Replicate 2 0.6385 0.734 0.1295 0.6155 0.773 0.666 0.665 0.729 0.085
Colony 2, Replicate 3 0.6445 0.731 0.131 0.6445 0.754 0.611 0.6975 0.7275 0.085
Colony 2, Replicate 4 0.626 0.668 0.128 0.67 0.759 0.6395 0.6885 0.695 0.085

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)
Colony 1, Replicate 1 32 79 255 113 30 145 28 26 25
Colony 1, Replicate 2 36 81 225 130 25 129 35 37 23
Colony 1, Replicate 3 32 82 278 122 29 133 29 29 29
Colony 1, Replicate 4 27 88 258 112 31 134 33 27 20
Colony 2, Replicate 1 30 48 255 114 36 30 27 25 22
Colony 2, Replicate 2 36 66 253 115 29 41 40 27 17
Colony 2, Replicate 3 36 71 255 110 30 35 29 33 34
Colony 2, Replicate 4 34 62 249 118 36 28 40 41 33

Hour 2

Hour 2: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6 LB + Chlor (blank)
Colony 1, Replicate 1 42 385 338 391 31 469 152 38 15
Colony 1, Replicate 2 37 345 322 459 28 403 146 35 12
Colony 1, Replicate 3 25 350 323 406 25 474 126 37 25
Colony 1, Replicate 4 36 327 352 417 38 449 118 33 17
Colony 2, Replicate 1 32 304 315 352 37 155 130 25 37
Colony 2, Replicate 2 26 297 319 388 39 156 116 45 47
Colony 2, Replicate 3 32 273 270 370 48 125 92 37 24
Colony 2, Replicate 4 34 277 269 381 54 128 111 33 40

Hour 4

Hour 4: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6 LB + Chlor (blank)
Colony 1, Replicate 1 31 783 497 893 40 1245 188 36 42
Colony 1, Replicate 2 30 730 495 825 34 801 183 39 35
Colony 1, Replicate 3 61 719 494 792 46 931 178 43 39
Colony 1, Replicate 4 35 746 523 832 39 829 185 42 25
Colony 2, Replicate 1 36 657 392 853 44 286 182 54 12
Colony 2, Replicate 2 27 649 324 821 59 342 176 46 25
Colony 2, Replicate 3 39 645 365 701 58 350 208 52 28
Colony 2, Replicate 4 38 666 363 832 52 356 241 47 26

Hour 6

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 27 1181 873 1539 46 1396 280 59 47
Colony 1, Replicate 2 43 1108 635 1463 51 1281 264 45 25
Colony 1, Replicate 3 26 1115 625 1454 65 1287 151 25 24
Colony 1, Replicate 4 52 1141 614 1422 72 1252 262 28 30
Colony 2, Replicate 1 35 1360 651 1352 55 632 316 41 24
Colony 2, Replicate 2 44 1091 381 1176 57 581 233 33 22
Colony 2, Replicate 3 40 1065 373 1299 50 546 248 31 39
Colony 2, Replicate 4 33 1034 376 1363 49 583 252 47 14