InterLab
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
- 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
- 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.)
- 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
Method:
Prepare the fluorescein stock solution:
- Spin down fluorescein stock tube to make sure pellet is at the bottom of tube
Prepare the serial dilutions of fluorescein:
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 |
Requirements
- 2L Erlenmeyer flask
- LB broth powder
- Bacteriological Agar
- Distilled or milliQ water
- Magnetic stirrer and bead(optional)
- Measuring cylinder
- Autoclave tape
- Cotton plug(for the flask)
- Aluminium foil
- Autoclaving machine
- Petri plates
Procedure
- Prepare 1L of LB liquid (25g broth powder per 1L) in a 2000mL Erlenmeyer flask.
- Add 15g (1.5%) bacteriological agar
- Use magnetic stirrer and bead to mix , if not available simply swirl to mix. Powder will not dissolve completely.
- Make a nice cotton plug by rolling and folding a big piece of cotton and put it on the mouth of the flask(Try pulling it out once, if it makes a noise then, Bravo!)
- Add a fresh piece of aluminum foil to cover the top, and add a fresh piece of autoclave tape.
- Autoclave at 121°C
- After cycle is complete, remove to benchtop to cool to ~50 C. (As a ballpark, this is still quite warm, but cool enough to leave your gloved hand on the outside of the flask for several seconds without burning it. The media should still be liquid but should be just beginning to thicken)
- If you are adding antibiotics, add them at this point, and swirl the flask vigorously for several seconds to mix (1000 times dilution of the stock solution)
- Set out your plates, and label the bottom with the appropriate antibiotic: A=ampicillin, C=chloramphenicol, K=kanamycin, T=tetracycline, S=streptomycin
- Pour out the media into petri dishes as follows: Remove the lid to a dish, and remove the foil cover to the flask. Pour just enough LB agar into the petri dish to completely cover the bottom of the dish, then replace the lid.
- Finish pouring dishes within 5 minutes of adding the antibiotic.
- Dishes should be stored UPSIDE DOWN in the cold room.
Requirements
- 1L autoclave bottle or erlenmeyer flask
- 6.25g LB broth powder
- 2.5mM KCl
- 20mM MgSO 4 .7H 2 O
- Distilled or milliQ water
- Magnetic stirrer and bead(optional)
- Measuring cylinder
- Autoclave tape
- Cotton plug(for the flask)
- Aluminium foil
- Autoclaving machine
Procedure
- In a 1L autoclave bottle or flask , add 6.25g LB broth powder.
- Calculate and weigh 2.5mM KCl and 20mM MgSO 4 .7H 2 O.
- Adjust pH to 7.5(using NaOH)
- Add distilled water to make the solution upto 250mL.
- Either Swirl or use magnetic stirrer and a bead to mix(might not dissolve completely) .
- Make a nice cotton plug by rolling and folding a big piece of cotton and put it on the mouth of the flask.(Try pulling it out once, if it makes a noise then, Bravo!)
- Put an Aluminium foil on top of it and a piece of autoclave tape
- Keep the cap slightly open if using a bottle.
- Autoclave at 121°C
- Cool to RT before use. Do not tighten cap until cool.
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