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<h3>OD600 Reference Point</h3> | <h3>OD600 Reference Point</h3> | ||
+ | <h5> 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 <em>E. coli</em> DH5 alpha cells | ||
{{Data/csv/OD600_reference_point}} | {{Data/csv/OD600_reference_point}} | ||
</div> | </div> |
Revision as of 18:14, 19 July 2017
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
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:
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 |