Template:Virginia/InterLab

InterLab

We did this year's iGEM InterLab study!
Our original Excel file can be downloaded here.
Here is the protocol that we followed.

OD600 Reference Point - LUDOX Protocol

LUDOX CL-X H2O
Replicate 1 0.113 0.087
Replicate 2 0.105 0.085
Replicate 3 0.119 0.084
Replicate 4 0.103 0.085
Arith. Mean 0.110 0.085
Corrected Abs600 0.025
Reference OD600 0.063
OD600/Abs600 2.545

Particle Standard Curve: Microsphere Protocol

Need to color code but: Abs600 measurements are blue cells. Gold cells are calculated.






Number of Particles 2.35E+08 1.18E+08 5.88E+07 2.94E+07 1.47E+07 7.35E+06 3.68E+06 1.84E+06 9.19E+05 4.60E+05 2.30E+05 0
Replicate 1 0.534 0.337 0.211 0.148 0.124 0.102 0.094 0.091 0.089 0.089 0.088 0.087
Replicate 2 0.538 0.329 0.204 0.145 0.117 0.102 0.099 0.092 0.089 0.089 0.086 0.085
Replicate 3 0.543 0.332 0.209 0.145 0.116 0.107 0.093 0.090 0.088 0.089 0.086 0.083
Replicate 4 0.545 0.314 0.208 0.143 0.119 0.102 0.092 0.089 0.089 0.089 0.086 0.083
Arith. Mean 0.540 0.328 0.208 0.145 0.119 0.103 0.095 0.091 0.089 0.089 0.087 0.085
Arith. Std.Dev. 0.005 0.010 0.003 0.002 0.004 0.003 0.003 0.001 0.001 0.000 0.001 0.002
Arith. Net Mean 0.456 0.244 0.124 0.061 0.035 0.019 0.010 0.006 0.004 0.004 0.002

<img src="T--Virginia--2018_ParticleStandardCurve.png" alt="ParticleStandardCurve"> <img src="T--Virginia--2018_ParticleStandardCurve_log.png" alt="ParticleStandardCurve_log">

Particles/OD

Note: Final scaling level determined from medium-high points likely to be less impacted by saturation or pipetting error. If needed, you can shift which points are used, but it is likely better to correct instrument settings and protocol.


Number of Particles 2.35E+08 1.18E+08 5.88E+07 2.94E+07 1.47E+07 7.35E+06 3.68E+06 1.84E+06 9.19E+05 4.60E+05 2.30E+05
Mean particles / Abs600 5.17E+08 4.83E+08 4.76E+08 4.84E+08 4.26E+08 3.92E+08 3.68E+08 3.06E+08 2.16E+08 1.02E+08 1.15E+08
Mean of med-high levels: 4.52E+08

Cospheric Monodisperse Silica Microspheres 0.961um diameter


Spheres/gram Grams/mL Spheres/0.55mL Resuspend volume mL: Dilution X: Total volume mL: Particles/mL: Well volume (mL) Initial particles:
1.20E+12 1.8 1.19E+12 2.55 100 255 4.71E+09 0.1 4.71E+08


Raw Plate Reader Measurements

Fluorescence Raw Readings:












Hour 0: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6 LB + Chlor (blank)
Colony 1, Replicate 1 124.47 172.15 136.79 137.65 115.66 132.36 127.35 135.34 112.83
Colony 1, Replicate 2 121.01 142.23 113.72 130.07 93.041 106.03 104.61 113.2 98.307
Colony 1, Replicate 3 123.38 148.37 112.73 131.37 90.708 106.35 104.49 113.95 95.141
Colony 1, Replicate 4 119.97 141.57 113.49 124.8 93.35 107.29 102.57 111.37 96.012
Colony 2, Replicate 1 121.32 134.7 116.21 169.04 93.865 110.22 152.05 111.67 99.513
Colony 2, Replicate 2 134.51 141.18 115.5 131.01 94.94 107.69 124.91 114.24 96.457
Colony 2, Replicate 3 130.47 141.41 114.54 130.7 92.262 111.95 128.96 115.03 98.671
Colony 2, Replicate 4 129.77 160.97 142.19 130.7 112.12 128.64 155.08 129.88 122.04









Hour 6: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6 LB + Chlor (blank)
Colony 1, Replicate 1 147.58 923.47 481.57 768.79 145.84 158.29 196.74 472.33 130.21
Colony 1, Replicate 2 135.57 868.37 443.8 771.52 133.54 150.48 174.77 442.69 108.87
Colony 1, Replicate 3 133.91 821.45 435.09 749.58 135.07 147.14 176.84 433.8 106.07
Colony 1, Replicate 4 131.16 825.34 424.75 702.31 136.56 145.48 178.85 435.43 101.34
Colony 2, Replicate 1 129.25 736.73 506.04 721.65 130.79 152.77 263.05 404.78 105.13
Colony 2, Replicate 2 129.93 746.95 522.99 752.02 130.12 154.08 274.44 99.78 102.2
Colony 2, Replicate 3 134.9 737.42 519.61 760.98 129.32 162.53 277.69 390.5 104.1
Colony 2, Replicate 4 136.44 824.81 585.7 749.59 145.38 171.03 319.39 429.11 122.55

Abs600 Raw Readings















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 0.114 0.112 0.112 0.116 0.115 0.113 0.11 0.112 0.097
Colony 1, Replicate 2 0.113 0.112 0.108 0.108 0.109 0.109 0.106 0.112 0.097
Colony 1, Replicate 3 0.114 0.11 0.106 0.108 0.109 0.108 0.106 0.111 0.093
Colony 1, Replicate 4 0.114 0.11 0.106 0.108 0.109 0.107 0.105 0.109 0.095
Colony 2, Replicate 1 0.115 0.114 0.108 0.123 0.111 0.112 0.124 0.114 0.094
Colony 2, Replicate 2 0.115 0.112 0.107 0.108 0.109 0.109 0.11 0.111 0.096
Colony 2, Replicate 3 0.114 0.111 0.107 0.107 0.11 0.111 0.116 0.111 0.093
Colony 2, Replicate 4 0.115 0.113 0.109 0.11 0.11 0.109 0.11 0.11 0.095








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 0.585 0.526 0.49 0.538 0.594 0.545 0.558 0.595 0.096
Colony 1, Replicate 2 0.563 0.533 0.483 0.533 0.582 0.544 0.541 0.56 0.093
Colony 1, Replicate 3 0.595 0.538 0.498 0.525 0.585 0.566 0.54 0.531 0.094
Colony 1, Replicate 4 0.586 0.532 0.49 0.494 0.581 0.571 0.5 0.569 0.094
Colony 2, Replicate 1 0.548 0.543 0.502 0.527 0.553 0.565 0.544 0.564 0.094
Colony 2, Replicate 2 0.566 0.543 0.482 0.536 0.544 0.544 0.526 0.541 0.096
Colony 2, Replicate 3 0.528 0.514 0.484 0.535 0.55 0.519 0.507 0.561 0.093
Colony 2, Replicate 4 0.52 0.516 0.462 0.496 0.53 0.523 0.488 0.524 0.097


Fluorescence per OD

uM Fluorescein / OD







Hour 0: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6
Colony 1, Replicate 1 0.066 0.382 0.154 0.126 0.015 0.118 0.108 0.145
Colony 1, Replicate 2 0.137 0.283 0.135 0.279 -0.042 0.062 0.068 0.096
Colony 1, Replicate 3 0.130 0.302 0.131 0.233 -0.027 0.072 0.069 0.101
Colony 1, Replicate 4 0.122 0.293 0.153 0.214 -0.018 0.091 0.063 0.106
Colony 2, Replicate 1 0.100 0.170 0.115 0.231 -0.032 0.057 0.169 0.059
Colony 2, Replicate 2 0.193 0.270 0.167 0.278 -0.011 0.083 0.196 0.114
Colony 2, Replicate 3 0.146 0.229 0.109 0.221 -0.036 0.071 0.127 0.088
Colony 2, Replicate 4 0.037 0.209 0.139 0.056 -0.064 0.046 0.213 0.050


























































Hour 6: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6
Colony 1, Replicate 1 0.003 0.178 0.086 0.139 0.003 0.006 0.014 0.066
Colony 1, Replicate 2 0.005 0.167 0.083 0.145 0.005 0.009 0.014 0.069
Colony 1, Replicate 3 0.005 0.156 0.079 0.144 0.006 0.008 0.015 0.072
Colony 1, Replicate 4 0.006 0.160 0.079 0.145 0.007 0.009 0.018 0.068
Colony 2, Replicate 1 0.005 0.136 0.095 0.137 0.005 0.010 0.034 0.062
Colony 2, Replicate 2 0.006 0.139 0.105 0.143 0.006 0.011 0.039 0.065
Colony 2, Replicate 3 0.007 0.145 0.103 0.143 0.005 0.013 0.040 0.059
Colony 2, Replicate 4 0.003 0.162 0.123 0.152 0.005 0.011 0.049 0.069


Net Fluorescein a.u.












































Hour 0: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6
Colony 1, Replicate 1 11.64 59.32 23.96 24.82 2.83 19.53 14.52 22.51
Colony 1, Replicate 2 22.70 43.92 15.41 31.76 -5.27 7.72 6.30 14.89
Colony 1, Replicate 3 28.24 53.23 17.59 36.23 -4.43 11.21 9.35 18.81
Colony 1, Replicate 4 23.96 45.56 17.48 28.79 -2.66 11.28 6.56 15.36
Colony 2, Replicate 1 21.81 35.19 16.70 69.53 -5.65 10.71 52.54 12.16
Colony 2, Replicate 2 38.05 44.72 19.04 34.55 -1.52 11.23 28.45 17.78
Colony 2, Replicate 3 31.80 42.74 15.87 32.03 -6.41 13.28 30.29 16.36
Colony 2, Replicate 4 7.73 38.93 20.15 8.66 -9.92 6.60 33.04 7.84


























Hour 6: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6
Colony 1, Replicate 1 17.37 793.26 351.36 638.58 15.63 28.08 66.53 342.12
Colony 1, Replicate 2 26.70 759.50 334.93 662.65 24.67 41.61 65.90 333.82
Colony 1, Replicate 3 27.84 715.38 329.02 643.51 29.00 41.07 70.77 327.73
Colony 1, Replicate 4 29.82 724.00 323.41 600.97 35.22 44.14 77.51 334.09
Colony 2, Replicate 1 24.12 631.60 400.91 616.52 25.66 47.64 157.92 299.65
Colony 2, Replicate 2 27.73 644.75 420.79 649.82 27.92 51.88 172.24 297.58
Colony 2, Replicate 3 30.80 633.32 415.51 656.88 25.22 58.43 173.59 286.40
Colony 2, Replicate 4 13.89 702.26 463.15 627.04 22.83 48.48 196.84 306.56

Net Abs 600

























































Hour 0: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6
Colony 1, Replicate 1 0.017 0.015 0.015 0.019 0.018 0.016 0.013 0.015
Colony 1, Replicate 2 0.016 0.015 0.011 0.011 0.012 0.012 0.009 0.015
Colony 1, Replicate 3 0.021 0.017 0.013 0.015 0.016 0.015 0.013 0.018
Colony 1, Replicate 4 0.019 0.015 0.011 0.013 0.014 0.012 0.010 0.014
Colony 2, Replicate 1 0.021 0.020 0.014 0.029 0.017 0.018 0.030 0.020
Colony 2, Replicate 2 0.019 0.016 0.011 0.012 0.013 0.013 0.014 0.015
Colony 2, Replicate 3 0.021 0.018 0.014 0.014 0.017 0.018 0.023 0.018
Colony 2, Replicate 4 0.020 0.018 0.014 0.015 0.015 0.014 0.015 0.015
























































Hour 6: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6
Colony 1, Replicate 1 0.489 0.430 0.394 0.442 0.498 0.449 0.462 0.499
Colony 1, Replicate 2 0.470 0.440 0.390 0.440 0.489 0.451 0.448 0.467
Colony 1, Replicate 3 0.501 0.444 0.404 0.431 0.491 0.472 0.446 0.437
Colony 1, Replicate 4 0.492 0.438 0.396 0.400 0.487 0.477 0.406 0.475
Colony 2, Replicate 1 0.454 0.449 0.408 0.433 0.459 0.471 0.450 0.470
Colony 2, Replicate 2 0.470 0.447 0.386 0.440 0.448 0.448 0.430 0.445
Colony 2, Replicate 3 0.435 0.421 0.391 0.442 0.457 0.426 0.414 0.468
Colony 2, Replicate 4 0.423 0.419 0.365 0.399 0.433 0.426 0.391 0.427

Fluorescence per Particle

MEFL / particle


































Hour 0: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6
Colony 1, Replicate 1 2.24E+03 1.29E+04 5.23E+03 4.27E+03 5.14E+02 3.99E+03 3.65E+03 4.91E+03
Colony 1, Replicate 2 4.64E+03 9.58E+03 4.58E+03 9.45E+03 -1.44E+03 2.11E+03 2.29E+03 3.25E+03
Colony 1, Replicate 3 4.40E+03 1.02E+04 4.43E+03 7.90E+03 -9.06E+02 2.44E+03 2.35E+03 3.42E+03
Colony 1, Replicate 4 4.12E+03 9.94E+03 5.20E+03 7.24E+03 -6.22E+02 3.07E+03 2.15E+03 3.59E+03
Colony 2, Replicate 1 3.40E+03 5.76E+03 3.90E+03 7.84E+03 -1.09E+03 1.95E+03 5.73E+03 1.99E+03
Colony 2, Replicate 2 6.55E+03 9.14E+03 5.66E+03 9.42E+03 -3.82E+02 2.83E+03 6.65E+03 3.88E+03
Colony 2, Replicate 3 4.95E+03 7.77E+03 3.71E+03 7.48E+03 -1.23E+03 2.41E+03 4.31E+03 2.97E+03
Colony 2, Replicate 4 1.26E+03 7.08E+03 4.71E+03 1.89E+03 -2.16E+03 1.54E+03 7.21E+03 1.71E+03
























































Hour 6: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6
Colony 1, Replicate 1 1.16E+02 6.03E+03 2.92E+03 4.73E+03 1.03E+02 2.05E+02 4.71E+02 2.24E+03
Colony 1, Replicate 2 1.86E+02 5.65E+03 2.81E+03 4.93E+03 1.65E+02 3.02E+02 4.81E+02 2.34E+03
Colony 1, Replicate 3 1.82E+02 5.27E+03 2.66E+03 4.88E+03 1.93E+02 2.85E+02 5.19E+02 2.45E+03
Colony 1, Replicate 4 1.98E+02 5.41E+03 2.67E+03 4.91E+03 2.37E+02 3.03E+02 6.25E+02 2.30E+03
Colony 2, Replicate 1 1.74E+02 4.60E+03 3.21E+03 4.66E+03 1.83E+02 3.31E+02 1.15E+03 2.09E+03
Colony 2, Replicate 2 1.93E+02 4.72E+03 3.57E+03 4.83E+03 2.04E+02 3.79E+02 1.31E+03 2.19E+03
Colony 2, Replicate 3 2.32E+02 4.92E+03 3.48E+03 4.86E+03 1.81E+02 4.49E+02 1.37E+03 2.00E+03
Colony 2, Replicate 4 1.07E+02 5.48E+03 4.15E+03 5.14E+03 1.72E+02 3.72E+02 1.65E+03 2.35E+03

Net Fluorescein a.u.


















































Hour 0: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6
Colony 1, Replicate 1 11.64 59.32 23.96 24.82 2.83 19.53 14.52 22.51
Colony 1, Replicate 2 22.70 43.92 15.41 31.76 -5.27 7.72 6.30 14.89
Colony 1, Replicate 3 28.24 53.23 17.59 36.23 -4.43 11.21 9.35 18.81
Colony 1, Replicate 4 23.96 45.56 17.48 28.79 -2.66 11.28 6.56 15.36
Colony 2, Replicate 1 21.81 35.19 16.70 69.53 -5.65 10.71 52.54 12.16
Colony 2, Replicate 2 38.05 44.72 19.04 34.55 -1.52 11.23 28.45 17.78
Colony 2, Replicate 3 31.80 42.74 15.87 32.03 -6.41 13.28 30.29 16.36
Colony 2, Replicate 4 7.73 38.93 20.15 8.66 -9.92 6.60 33.04 7.84



























Hour 6: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6
Colony 1, Replicate 1 17.37 793.26 351.36 638.58 15.63 28.08 66.53 342.12
Colony 1, Replicate 2 26.70 759.50 334.93 662.65 24.67 41.61 65.90 333.82
Colony 1, Replicate 3 27.84 715.38 329.02 643.51 29.00 41.07 70.77 327.73
Colony 1, Replicate 4 29.82 724.00 323.41 600.97 35.22 44.14 77.51 334.09
Colony 2, Replicate 1 24.12 631.60 400.91 616.52 25.66 47.64 157.92 299.65
Colony 2, Replicate 2 27.73 644.75 420.79 649.82 27.92 51.88 172.24 297.58
Colony 2, Replicate 3 30.80 633.32 415.51 656.88 25.22 58.43 173.59 286.40
Colony 2, Replicate 4 13.89 702.26 463.15 627.04 22.83 48.48 196.84 306.56

Net Abs 600

























































Hour 0: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6
Colony 1, Replicate 1 0.017 0.015 0.015 0.019 0.018 0.016 0.013 0.015
Colony 1, Replicate 2 0.016 0.015 0.011 0.011 0.012 0.012 0.009 0.015
Colony 1, Replicate 3 0.021 0.017 0.013 0.015 0.016 0.015 0.013 0.018
Colony 1, Replicate 4 0.019 0.015 0.011 0.013 0.014 0.012 0.010 0.014
Colony 2, Replicate 1 0.021 0.020 0.014 0.029 0.017 0.018 0.030 0.020
Colony 2, Replicate 2 0.019 0.016 0.011 0.012 0.013 0.013 0.014 0.015
Colony 2, Replicate 3 0.021 0.018 0.014 0.014 0.017 0.018 0.023 0.018
Colony 2, Replicate 4 0.020 0.018 0.014 0.015 0.015 0.014 0.015 0.015
























































Hour 6: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6
Colony 1, Replicate 1 0.489 0.430 0.394 0.442 0.498 0.449 0.462 0.499
Colony 1, Replicate 2 0.470 0.440 0.390 0.440 0.489 0.451 0.448 0.467
Colony 1, Replicate 3 0.501 0.444 0.404 0.431 0.491 0.472 0.446 0.437
Colony 1, Replicate 4 0.492 0.438 0.396 0.400 0.487 0.477 0.406 0.475
Colony 2, Replicate 1 0.454 0.449 0.408 0.433 0.459 0.471 0.450 0.470
Colony 2, Replicate 2 0.470 0.447 0.386 0.440 0.448 0.448 0.430 0.445
Colony 2, Replicate 3 0.435 0.421 0.391 0.442 0.457 0.426 0.414 0.468
Colony 2, Replicate 4 0.423 0.419 0.365 0.399 0.433 0.426 0.391 0.427