Team:IISER-Pune-India/InterLab

Interlab Studies

InterLab

Interlab studies is a great initiative taken by the iGEM headquaters. Reproducibilty is a very big problem in biology. Repeating a result is not always a simple task.Repeating the research can both correct inaccuracies and deepen our understanding of our work.

Here is the protocol that we followed and our results:

Calibration Experiments

Materials:

  • 1ml LUDOX (provided in kit)
  • H20 (provided by team)
  • 96 well plate, black with flat, transparent/clear bottom preferred (provided by team)

Method

  • Add 100 µl LUDOX into wells A1, B1, C1, D1 (or 1 mL LUDOX into cuvette)
  • Add 100 µl of H2O into wells A2, B2, C2, D2 (or 1 mL H2O into cuvette)
  • Measure absorbance 600 nm of all samples in all standard measurement modes in instrument
  • Record the data in the table below or in your notebook
  • Import data into Excel (OD600 reference point tab)​ Sheet_1 provided

Materials:

  • fluorescein (provided in kit)
  • 10ml 1xPBS (phosphate buffered saline; provided by team)
  • Corning 96 well plate, black,flat bottom,half-well

Method:

Prepare the fluorescein stock solution:

  • Spin down fluorescein stock tube to make sure pellet is at the bottom of tube
  • Prepare 2x fluorescein stock solution (100 µM) by resuspending fluorescein in 1 mL of 1xPBS. [Note​: it is important that the fluorescein is properly dissolved. To check this, after the resuspension you should pipette up and down and examine the solution in the pipette tip – if any particulates are visible in the pipette tip continue to mix the solution until they disappear.
  • Dilute the 2x fluorescein stock solution with 1xPBS to make a 1x fluorescein solution and resulting concentration of fluorescein stock solution 50 µM (500µL of 2x fluorescein in 500 µL 1x PBS will make 1 mL of 50 µM (1x) fluorescein solution.)

Prepare the serial dilutions of fluorescein:

  • Add 100 µl of PBS​ into wells A2, B2, C2, D2....A12, B12, C12, D12
  • Add 200 µl​ of fluorescein 1x stock​ solution into A1, B1, C1, D1
  • Transfer 100 µl of fluorescein stock solution from A1 into A2.
  • Mix A2 by pipetting up and down 3x and transfer 100 µl into A3…
  • Mix A3 by pipetting up and down 3x and transfer 100 µl into A4...
  • Mix A4 by pipetting up and down 3x and transfer 100 µl into A5...
  • Mix A5 by pipetting up and down 3x and transfer 100 µl into A6...
  • Mix A6 by pipetting up and down 3x and transfer 100 µl into A7...
  • Mix A7 by pipetting up and down 3x and transfer 100 µl into A8...
  • Mix A8 by pipetting up and down 3x and transfer 100 µl into A9...
  • Mix A9 by pipetting up and down 3x and transfer 100 µl into A10...
  • Mix A10 by pipetting up and down 3x and transfer 100 µl into A11...
  • Mix A11 by pipetting up and down 3x and transfer 100 µl into liquid waste
  • Repeat dilution series for rows B, C
  • Measure fluorescence of all samples in all standard measurement modes in instrumen
  • Record the data in your noteboo
  • Import data into Excel (fluorescein standard curve tab)​ Sheet_1 provided

Fluorescence measurement were taken using Thermo Scientific Varioskan Flash plate reader.

  • Plate reader settings:
  • Light source: Xenon flash lamp
  • Excitation wavelength-480nm
  • Emission wavelength-510nm
  • Measurement time-100ms
  • Excitation bandwidth-5nm
  • Emission bandwidth-12nm
Cell Meaurement Protocol

Materials:

  • Competent cells (Escherichia coli strain DH5α)
  • LB (Luria Bertani) media
  • Chloramphenicol (stock concentration 25 mg/mL dissolved in EtOH - working stock 25 ug/mL)
  • 50 ml Falcon tube (or equivalent, preferably amber or covered in foil to block light)
  • Incubator at 37°C
  • 1.5 ml eppendorf tubes for sample storage
  • Ice bucket with ice
  • Pipettes
  • Corning 96 well plate,transparent,full well,flat bottom
  • Corning 96 well plate,black,half well,flat bottom

    • Devices (from InterLab Measurement Kit):

    • Positive control
    • Negative control
    • Test Device 1: J23101+I13504
    • Test Device 2: J23106+I13504
    • Test Device 3: J23117+I13504
    • Test Device 4: J23101.BCD2.E0040.B0015
    • Test Device 5: J23106.BCD2.E0040.B0015
    • Test Device 6: J23117.BCD2.E0040.B0015

Method

Day 1​: transform Escherichia coli DH5α with these following plasmids:

  • Positive control
  • Negative control
  • Test Device 1: J23101+I13504
  • Test Device 2: J23106+I13504
  • Test Device 3: J23117+I13504
  • Test Device 4: J23101.BCD2.E0040.B0015
  • Test Device 5: J23106.BCD2.E0040.B0015
  • Test Device 6: J23117.BCD2.E0040.B0015

Day 2​: Pick 2 colonies from each of plate and inoculate it on 5mL LB medium + Chloramphenicol. Grow the cells overnight (16-18 hours) at 37°C and 220 rpm.

Day 3​: Cell growth, sampling, and assay

  • Set your instrument to read OD600 (as OD calibration setting)
  • Measure OD600 of the overnight cultures
  • Record data in your notebook
  • Import data into Excel (Dilution Calculation​)
  • Dilute the cultures to a target OD600 of 0.02 in 12 m​l LB medium + Chloramphenicol in 50 mL falcon tube (covered with foil to block light).
  • Incubate the cultures at 37°C and 220 rpm.
  • Take 500 µL samples of the cultures at 0, 2, 4, and 6 hours of incubation. (At each time point, you will take a sample from each of the 8 devices, two colonies per device, for a total of 16 samples per time point)
  • Place samples on ice or 4℃ fridge
  • At the end of sampling point you need to measure your samples (OD and Fl measurement)
  • Take two different plates(black and transparent), load 100µL of sample on black plate for fluorescence measurement and 120µL in transparent plate for OD measurement
  • Record data in your notebook
  • Import data into Excel
Raw Data for Fluorescein Std Curve
uM Fluorescein 50.00 25 12.5 6.25 3.125 1.5625 0.78125 0.390625 0.195313 0.097656 0.048828 0
Replicate 1 13414.1 11455.1 8410.2 5255.2 2576.53 1581.79 674.3 364.496 219.688 110.455 42.6915 0.116654
Replicate 2 13484.6 11463.9 8163.66 4963.2 2863.6 1566.24 705.598 438.018 189.537 88.2177 47.6698 0.181143
Replicate 3 13468.1 11541.2 8193.38 5177.1 2939.83 1587.29 823.477 422.591 233.03 115.595 54.3853 0.152354
Replicate 4 13574.4 9851.55 6476.66 5177.48 2949.9 1585.38 828.868 426.119 221.582 117.785 58.7254 0.159733
Arith. Mean 13485.3 11077.94 7810.975 5143.245 2832.465 1580.175 758.0608 412.806 215.9593 108.0132 50.868 0.152471
Arith. Std.Dev. 66.59474 818.5062 896.3049 125.5233 174.9193 9.565697 79.71022 32.87594 18.57476 13.54986 7.099149 0.026818
Raw Data for Cell Measurements
Experimental Values: Raw Abs600         Raw Fluorescence                                              
Sample set: Replicate 1 Replicate 2 Replicate 3 Replicate 4   Replicate 1 Replicate 2 Replicate 3 Replicate 4                                        
Blank media 0.039 0.038 0.038 0.038   11.92 11.32 11.99 11.91                                        
Blank mean: 0.03825         11.785         OD - Background         Fluorescence - Background         uM Fluorescein / OD600         Summary Statistics      
Hour 0:                     Replicate 1 Replicate 2 Replicate 3 Replicate 4   Replicate 1 Replicate 2 Replicate 3 Replicate 4   Replicate 1 Replicate 2 Replicate 3 Replicate 4   Arith. Mean Arith. Std.Dev. Geo. Mean Geo. Std. Dev.
Negative Control (Colony 1) 0.047 0.047 0.047 0.047   12.75 12.83 13.35 12.82   0.00875 0.00875 0.00875 0.00875   0.965 1.045 1.565 1.035   0.10 0.10 0.16 0.10   0.12 0.03 0.11 1.246
Negative Control (Colony 2) 0.047 0.047 0.047 0.046   10.89 9.781 7.717 12.44   0.00875 0.00875 0.00875 0.00775   -0.895 -2.004 -4.068 0.655   -0.09 -0.20 -0.41 0.07   -0.16 0.20 #NUM! #NUM!
Positive Control (Colony 1) 0.047 0.048 0.047 0.047   12.13 12.44 13.99 13.51   0.00875 0.00975 0.00875 0.00875   0.345 0.655 2.205 1.725   0.03 0.06 0.22 0.17   0.12 0.09 0.09 2.410
Positive Control (Colony 2) 0.049 0.047 0.046 0.046   13.46 12.61 13.06 13.77   0.01075 0.00875 0.00775 0.00775   1.675 0.825 1.275 1.985   0.14 0.08 0.14 0.22   0.15 0.06 0.14 1.505
Test Device 1: J23101.BCD2.E0040.B0015 (Colony 1) 0.049 0.05 0.048 0.049   18.19 20.69 19.15 19.37   0.01075 0.01175 0.00975 0.01075   6.405 8.905 7.365 7.585   0.52 0.66 0.66 0.62   0.62 0.07 0.61 1.119
Test Device 1: J23101.BCD2.E0040.B0015 (Colony 2) 0.047 0.046 0.047 0.046   21.62 19.76 20.76 18.28   0.00875 0.00775 0.00875 0.00775   9.835 7.975 8.975 6.495   0.99 0.90 0.90 0.73   0.88 0.10 0.88 1.132
Test Device 2: J23106.BCD2.E0040.B0015 (Colony 1) 0.047 0.046 0.047 0.047   13.95 14.37 14.62 14.54   0.00875 0.00775 0.00875 0.00875   2.165 2.585 2.835 2.755   0.22 0.29 0.28 0.28   0.27 0.03 0.27 1.147
Test Device 2: J23106.BCD2.E0040.B0015 (Colony 2) 0.048 0.047 0.046 0.047   14.38 16.27 13.38 15.53   0.00975 0.00875 0.00775 0.00875   2.595 4.485 1.595 3.745   0.23 0.45 0.18 0.38   0.31 0.12 0.29 1.523
Test Device 3: J23117.BCD2.E0040.B0015 (Colony 1) 0.048 0.048 0.071 0.049   11.44 12.69 14.09 11.64   0.00975 0.00975 0.03275 0.01075   -0.345 0.905 2.305 -0.145   -0.03 0.08 0.06 -0.01   0.03 0.05 #NUM! #NUM!
Test Device 3: J23117.BCD2.E0040.B0015 (Colony 2) 0.047 0.046 0.045 0.051   13.57 13.91 12.65 13.24   0.00875 0.00775 0.00675 0.01275   1.785 2.125 0.865 1.455   0.18 0.24 0.11 0.10   0.16 0.06 0.15 1.504
Test Device 4: J23101+I13504 (Colony 1) 0.045 0.046 0.047 0.047   14.09 13.1 14.81 13.72   0.00675 0.00775 0.00875 0.00875   2.305 1.315 3.025 1.935   0.30 0.15 0.30 0.19   0.24 0.08 0.23 1.416
Test Device 4: J23101+I13504 (Colony 2) 0.048 0.046 0.046 0.046   15.34 14.29 13.91 13.42   0.00975 0.00775 0.00775 0.00775   3.555 2.505 2.125 1.635   0.32 0.28 0.24 0.18   0.26 0.06 0.25 1.267
Test Device 5: J23106+I13504 (Colony 1) 0.046 0.046 0.047 0.046   13.8 11.54 12.31 10.5   0.00775 0.00775 0.00875 0.00775   2.015 -0.245 0.525 -1.285   0.23 -0.03 0.05 -0.15   0.03 0.16 #NUM! #NUM!
Test Device 5: J23106+I13504 (Colony 2) 0.047 0.047 0.046 0.046   13.06 14.03 13.71 13.77   0.00875 0.00875 0.00775 0.00775   1.275 2.245 1.925 1.985   0.13 0.22 0.22 0.22   0.20 0.05 0.19 1.320
Test Device 6: J23117+I13504 (Colony 1) 0.046 0.047 0.047 0.047   13.49 12.07 12.23 12.83   0.00775 0.00875 0.00875 0.00875   1.705 0.285 0.445 1.045   0.19 0.03 0.04 0.10   0.09 0.07 0.07 2.353
Test Device 6: J23117+I13504 (Colony 2) 0.046 0.044 0.044 0.044   13.24 12.34 13.56 13.93   0.00775 0.00575 0.00575 0.00575   1.455 0.555 1.775 2.145   0.16 0.08 0.27 0.33   0.21 0.11 0.19 1.829
                                                           
Hour 2:                                                          
Negative Control (Colony 1) 0.055 0.068 0.054 0.053   13.36 12.83 12.3 12.85   0.01675 0.02975 0.01575 0.01475   1.575 1.045 0.515 1.065   0.08 0.03 0.03 0.06   0.05 0.03 0.05 1.690
Negative Control (Colony 2) 0.232 0.209 0.148 0.054   13.54 12.85 13.88 11.16   0.19375 0.17075 0.10975 0.01575   1.755 1.065 2.095 -0.625   0.01 0.01 0.02 -0.03   0.00 0.02 #NUM! #NUM!
Positive Control (Colony 1) 0.058 0.053 0.053 0.052   20.39 19.68 20.97 21   0.01975 0.01475 0.01475 0.01375   8.605 7.895 9.185 9.215   0.38 0.47 0.55 0.59   0.50 0.09 0.49 1.210
Positive Control (Colony 2) 0.152 0.104 0.053 0.054   21.64 20.63 19.6 20.02   0.11375 0.06575 0.01475 0.01575   9.855 8.845 7.815 8.235   0.08 0.12 0.46 0.46   0.28 0.21 0.21 2.540
Test Device 1: J23101.BCD2.E0040.B0015 (Colony 1) 0.051 0.054 0.049 0.049   26.75 29.96 29.37 29.77   0.01275 0.01575 0.01075 0.01075   14.965 18.175 17.585 17.985   1.03 1.01 1.43 1.47   1.24 0.25 1.22 1.226
Test Device 1: J23101.BCD2.E0040.B0015 (Colony 2) 0.204 0.193 0.195 0.136   29.84 32.35 33.21 26.81   0.16575 0.15475 0.15675 0.09775   18.055 20.565 21.425 15.025   0.10 0.12 0.12 0.13   0.12 0.02 0.12 1.154
Test Device 2: J23106.BCD2.E0040.B0015 (Colony 1) 0.052 0.05 0.05 0.05   31.21 29.42 29.86 31.01   0.01375 0.01175 0.01175 0.01175   19.425 17.635 18.075 19.225   1.24 1.32 1.35 1.43   1.33 0.08 1.33 1.063
Test Device 2: J23106.BCD2.E0040.B0015 (Colony 2) 0.054 0.076 0.053 0.052   30.14 35.8 34.28 31.55   0.01575 0.03775 0.01475 0.01375   18.355 24.015 22.495 19.765   1.02 0.56 1.34 1.26   1.04 0.35 0.99 1.491
Test Device 3: J23117.BCD2.E0040.B0015 (Colony 1) 0.054 0.055 0.057 0.054   14.69 14.47 16.36 16.08   0.01575 0.01675 0.01875 0.01575   2.905 2.685 4.575 4.295   0.16 0.14 0.21 0.24   0.19 0.05 0.18 1.278
Test Device 3: J23117.BCD2.E0040.B0015 (Colony 2) 0.055 0.054 0.053 0.052   17.35 16.78 18.05 15.31   0.01675 0.01575 0.01475 0.01375   5.565 4.995 6.265 3.525   0.29 0.28 0.37 0.22   0.29 0.06 0.29 1.230
Test Device 4: J23101+I13504 (Colony 1) 0.054 0.053 0.052 0.052   21.9 22.44 23.34 23.84   0.01575 0.01475 0.01375 0.01375   10.115 10.655 11.555 12.055   0.56 0.63 0.74 0.77   0.68 0.09 0.67 1.154
Test Device 4: J23101+I13504 (Colony 2) 0.051 0.052 0.051 0.05   25.2 25.93 25.8 24.39   0.01275 0.01375 0.01275 0.01175   13.415 14.145 14.015 12.605   0.92 0.90 0.96 0.94   0.93 0.03 0.93 1.029
Test Device 5: J23106+I13504 (Colony 1) 0.055 0.053 0.051 0.053   22.94 22.74 21.48 21.35   0.01675 0.01475 0.01275 0.01475   11.155 10.955 9.695 9.565   0.58 0.65 0.67 0.57   0.62 0.05 0.62 1.082
Test Device 5: J23106+I13504 (Colony 2) 0.057 0.056 0.055 0.055   22.02 22.25 22.51 21.09   0.01875 0.01775 0.01675 0.01675   10.235 10.465 10.725 9.305   0.48 0.52 0.56 0.49   0.51 0.04 0.51 1.075
Test Device 6: J23117+I13504 (Colony 1) 0.053 0.054 0.056 0.055   16.19 15.64 16.24 16.07   0.01475 0.01575 0.01775 0.01675   4.405 3.855 4.455 4.285   0.26 0.21 0.22 0.22   0.23 0.02 0.23 1.094
Test Device 6: J23117+I13504 (Colony 2) 0.051 0.051 0.049 0.05   16.13 16.2 16.26 17.05   0.01275 0.01275 0.01075 0.01175   4.345 4.415 4.475 5.265   0.30 0.30 0.36 0.39   0.34 0.05 0.34 1.145
                                                           
Hour 4:                                                          
Negative Control (Colony 1) 0.191 0.15 0.134 0.139   14.45 15.48 14.63 13.73   0.15275 0.11175 0.09575 0.10075   2.665 3.695 2.845 1.945   0.02 0.03 0.03 0.02   0.02 0.01 0.02 1.370
Negative Control (Colony 2) 0.147 0.161 0.151 0.142   14.89 15.77 16.32 15.74   0.10875 0.12275 0.11275 0.10375   3.105 3.985 4.535 3.955   0.03 0.03 0.04 0.03   0.03 0.00 0.03 1.169
Positive Control (Colony 1) 0.184 0.139 0.13 0.127   50.64 49.12 44.43 39.3   0.14575 0.10075 0.09175 0.08875   38.855 37.335 32.645 27.515   0.23 0.32 0.31 0.27   0.29 0.04 0.28 1.161
Positive Control (Colony 2) 0.157 0.14 0.133 0.134   58.28 57.94 52.46 52.37   0.11875 0.10175 0.09475 0.09575   46.495 46.155 40.675 40.585   0.34 0.40 0.38 0.37   0.37 0.02 0.37 1.063
Test Device 1: J23101.BCD2.E0040.B0015 (Colony 1) 0.057 0.056 0.055 0.054   48.17 41.93 44.44 43.61   0.01875 0.01775 0.01675 0.01575   36.385 30.145 32.655 31.825   1.70 1.49 1.71 1.77   1.67 0.12 1.66 1.079
Test Device 1: J23101.BCD2.E0040.B0015 (Colony 2) 0.056 0.054 0.054 0.052   46.69 50.03 47.27 45.78   0.01775 0.01575 0.01575 0.01375   34.905 38.245 35.485 33.995   1.72 2.13 1.97 2.17   2.00 0.20 1.99 1.110
Test Device 2: J23106.BCD2.E0040.B0015 (Colony 1) 0.143 0.13 0.129 0.116   76.83 75.98 72.69 70.29   0.10475 0.09175 0.09075 0.07775   65.045 64.195 60.905 58.505   0.54 0.61 0.59 0.66   0.60 0.05 0.60 1.084
Test Device 2: J23106.BCD2.E0040.B0015 (Colony 2) 0.131 0.106 0.1 0.096   75.71 70.61 69.71 75.01   0.09275 0.06775 0.06175 0.05775   63.925 58.825 57.925 63.225   0.60 0.76 0.82 0.96   0.79 0.15 0.78 1.213
Test Device 3: J23117.BCD2.E0040.B0015 (Colony 1) 0.208 0.152 0.144 0.126   18.56 17.22 16.66 15.38   0.16975 0.11375 0.10575 0.08775   6.775 5.435 4.875 3.595   0.03 0.04 0.04 0.04   0.04 0.00 0.04 1.092
Test Device 3: J23117.BCD2.E0040.B0015 (Colony 2) 0.155 0.131 0.114 0.115   16.97 16.11 16.05 17   0.11675 0.09275 0.07575 0.07675   5.185 4.325 4.265 5.215   0.04 0.04 0.05 0.06   0.05 0.01 0.05 1.214
Test Device 4: J23101+I13504 (Colony 1) 0.115 0.118 0.099 0.1   46.38 44.29 43.31 42.04   0.07675 0.07975 0.06075 0.06175   34.595 32.505 31.525 30.255   0.40 0.36 0.45 0.43   0.41 0.04 0.41 1.111
Test Device 4: J23101+I13504 (Colony 2) 0.107 0.103 0.099 0.094   46.3 44.96 44.81 42.31   0.06875 0.06475 0.06075 0.05575   34.515 33.175 33.025 30.525   0.44 0.45 0.48 0.48   0.46 0.02 0.46 1.044
Test Device 5: J23106+I13504 (Colony 1) 0.119 0.112 0.122 0.117   24.84 25.74 27.41 27.58   0.08075 0.07375 0.08375 0.07875   13.055 13.955 15.625 15.795   0.14 0.17 0.16 0.18   0.16 0.01 0.16 1.096
Test Device 5: J23106+I13504 (Colony 2) 0.139 0.125 0.147 0.135   24.09 22.96 26.29 26.06   0.10075 0.08675 0.10875 0.09675   12.305 11.175 14.505 14.275   0.11 0.11 0.12 0.13   0.12 0.01 0.12 1.083
Test Device 6: J23117+I13504 (Colony 1) 0.137 0.145 0.142 0.125   13.65 14.24 13.76 14.66   0.09875 0.10675 0.10375 0.08675   1.865 2.455 1.975 2.875   0.02 0.02 0.02 0.03   0.02 0.01 0.02 1.301
Test Device 6: J23117+I13504 (Colony 2) 0.11 0.111 0.11 0.116   14.42 14.08 13.93 14.6   0.07175 0.07275 0.07175 0.07775   2.635 2.295 2.145 2.815   0.03 0.03 0.03 0.03   0.03 0.00 0.03 1.107
                                                           
Hour 6:                                                          
Negative Control (Colony 1) 0.403 0.396 0.385 0.384   13.86 14.13 14.1 13.53   0.36475 0.35775 0.34675 0.34575   2.075 2.345 2.315 1.745   0.00 0.01 0.01 0.00   0.01 0.00 0.01 1.140
Negative Control (Colony 2) 0.282 0.399 0.446 0.312   13.67 13.87 14.02 13.94   0.24375 0.36075 0.40775 0.27375   1.885 2.085 2.235 2.155   0.01 0.01 0.00 0.01   0.01 0.00 0.01 1.209
Positive Control (Colony 1) 0.308 0.279 0.297 0.282   84.88 74.53 79.93 79.25   0.26975 0.24075 0.25875 0.24375   73.095 62.745 68.145 67.465   0.24 0.23 0.23 0.24   0.23 0.01 0.23 1.028
Positive Control (Colony 2) 0.393 0.303 0.237 0.411   79.7 68.25 81.04 81.93   0.35475 0.26475 0.19875 0.37275   67.915 56.465 69.255 70.145   0.17 0.19 0.31 0.16   0.21 0.07 0.20 1.336
Test Device 1: J23101.BCD2.E0040.B0015 (Colony 1) 0.069 0.059 0.068 0.057   53.36 48.94 53.32 52.33   0.03075 0.02075 0.02975 0.01875   41.575 37.155 41.535 40.545   1.19 1.57 1.22 1.90   1.47 0.33 1.44 1.248
Test Device 1: J23101.BCD2.E0040.B0015 (Colony 2) 0.07 0.069 0.068 0.068   54.14 55.06 56.59 55.34   0.03175 0.03075 0.02975 0.02975   42.355 43.275 44.805 43.555   1.17 1.23 1.32 1.28   1.25 0.07 1.25 1.054
Test Device 2: J23106.BCD2.E0040.B0015 (Colony 1) 0.308 0.307 0.288 0.324   146.1 129 142.7 146.6   0.26975 0.26875 0.24975 0.28575   134.315 117.215 130.915 134.815   0.44 0.38 0.46 0.41   0.42 0.03 0.42 1.082
Test Device 2: J23106.BCD2.E0040.B0015 (Colony 2) 0.403 0.427 0.217 0.287   125.9 120.6 132.6 121.6   0.36475 0.38875 0.17875 0.24875   114.115 108.815 120.815 109.815   0.27 0.25 0.59 0.39   0.37 0.16 0.35 1.487
Test Device 3: J23117.BCD2.E0040.B0015 (Colony 1) 0.398 0.399 0.392 0.343   16.24 15.33 14.79 16.13   0.35975 0.36075 0.35375 0.30475   4.455 3.545 3.005 4.345   0.01 0.01 0.01 0.01   0.01 0.00 0.01 1.261
Test Device 3: J23117.BCD2.E0040.B0015 (Colony 2) 0.3 0.295 0.209 0.181   15.95 15.66 15.93 15.44   0.26175 0.25675 0.17075 0.14275   4.165 3.875 4.145 3.655   0.01 0.01 0.02 0.02   0.02 0.00 0.02 1.318
Test Device 4: J23101+I13504 (Colony 1) 0.272 0.274 0.264 0.264   87.02 86.88 85.79 83.04   0.23375 0.23575 0.22575 0.22575   75.235 75.095 74.005 71.255   0.28 0.28 0.29 0.28   0.28 0.00 0.28 1.016
Test Device 4: J23101+I13504 (Colony 2) 0.296 0.236 0.225 0.228   97.47 97.03 97.78 82.43   0.25775 0.19775 0.18675 0.18975   85.685 85.245 85.995 70.645   0.29 0.38 0.40 0.33   0.35 0.05 0.35 1.158
Test Device 5: J23106+I13504 (Colony 1) 0.36 0.366 0.368 0.347   47.54 46.09 45.93 42.66   0.32175 0.32775 0.32975 0.30875   35.755 34.305 34.145 30.875   0.10 0.09 0.09 0.09   0.09 0.00 0.09 1.045
Test Device 5: J23106+I13504 (Colony 2) 0.327 0.324 0.294 0.309   36.82 41.73 41.66 37.26   0.28875 0.28575 0.25575 0.27075   25.035 29.945 29.875 25.475   0.08 0.09 0.10 0.08   0.09 0.01 0.09 1.138
Test Device 6: J23117+I13504 (Colony 1) 0.387 0.402 0.38 0.343   14.5 14.72 14.73 14.63   0.34875 0.36375 0.34175 0.30475   2.715 2.935 2.945 2.845   0.01 0.01 0.01 0.01   0.01 0.00 0.01 1.083
Test Device 6: J23117+I13504 (Colony 2) 0.351 0.374 0.356 0.375   14.53 14.82 14.32 14.82   0.31275 0.33575 0.31775 0.33675   2.745 3.035 2.535 3.035   0.01 0.01 0.01 0.01   0.01 0.00 0.01 1.060
OD600 Reference Point
LUDOX-HS4 H2O
Replicate 1 0.049 0.028
Replicate 2 0.054 0.026
Replicate 3 0.052 0.027
Replicate 4 0.058 0.028
Arith. Mean 0.05325 0.02725
Corrected Abs600 0.026
Reference OD600 0.0425
OD600/Abs600 1.6346154
Fluorescein Std Curves
Combined OD and Fluorescence Measurement Curves
These are the combined graphs of fluoroscence and OD measurements of all devices averaged for the two colonies-