Interlab
Introduction
Reliable and repeatable measurement is a key component of synthetic biology. However, it has been difficult to repeat the same measurements in different labs. There have been few opportunities to compare the fluorescence data because it has been reported in different units or because different groups process data in different ways.
With the aim to improve the measurement tools available to both the iGEM community and the synthetic biology community, the measurement committee invited all iGEM teams to participate in the fifth InterLab Study, which provides researchers with a detailed protocol and data analysis form, with the aim to produce common, comparable units for measuring GFP on different plate readers.
For a more detailed description visit the homepage of the iGEM's measurement committee. (https://2018.igem.org/Measurement/InterLab)
Device we received
| Positive Control (BBa_I20270) |
| Negative Control (BBa_R0040) |
| Test Device1 (BBa_J364000) |
| Test Device2 (BBa_J364001) |
| Test Device3 (BBa_J364002) |
| Test Device4 (BBa_J364007) |
| Test Device5 (BBa_J364008) |
| Test Device6 (BBa_J364009) |
Protocol
Strictly following the protocol provided by iGEM, we made three sets of unit calibration measurements: an OD600 reference point, a particle standard curve, and a fluorescein standard curve.
The transformation protocol(连接)
The interLab plate reader protocol（连接）
The raw data
| LUDOX CL-X | H2O | |
| Replicate 1 | 0.06 | 0.047 |
| Replicate 2 | 0.059 | 0.045 |
| Replicate 3 | 0.06 | 0.048 |
| Replicate 4 | 0.064 | 0.056 |
| Arith. Mean | 0.061 | 0.049 |
| Corrected Abs600 | 0.012 | |
| Reference OD600 | 0.063 | |
| OD600/Abs600 | 5.362 |
Stable 1. OD600Reference Point Stable 2. Particle Count
| 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 | 0.528 | 0.315 | 0.163 | 0.126 | 0.104 | 0.076 | 0.064 | 0.047 | 0.04 | 0.041 | 0.043 | 0.043 |
| Replicate 2 | 0.556 | 0.286 | 0.17 | 0.106 | 0.087 | 0.062 | 0.043 | 0.041 | 0.04 | 0.05 | 0.057 | 0.047 |
| Replicate 3 | 0.524 | 0.289 | 0.161 | 0.109 | 0.104 | 0.058 | 0.051 | 0.048 | 0.036 | 0.049 | 0.053 | 0.042 |
| Replicate 4 | 0.524 | 0.269 | 0.159 | 0.106 | 0.078 | 0.066 | 0.05 | 0.043 | 0.034 | 0.041 | 0.045 | 0.037 |
| Arith. Mean | 0.533 | 0.29 | 0.163 | 0.112 | 0.093 | 0.066 | 0.052 | 0.045 | 0.038 | 0.045 | 0.05 | 0.042 |
| Arith. Std.Dev. | 0.015 | 0.019 | 0.005 | 0.01 | 0.013 | 0.008 | 0.009 | 0.003 | 0.003 | 0.005 | 0.007 | 0.004 |
| Arith. Net Mean | 0.491 | 0.248 | 0.121 | 0.07 | 0.051 | 0.023 | 0.01 | 0.003 | -0.005 | 0.003 | 0.007 |
Stable 2. Particle Count
| Fluorescein uM | 10 | 5 | 2.5 | 1.25 | 0.625 | 0.313 | 0.156 | 0.078 | 0.039 | 0.0195 | 0.0098 | 0 |
| Replicate 1 | 18743 | 9954 | 5246 | 2864 | 1544 | 755 | 411 | 214 | 118 | 67 | 38 | 13 |
| Replicate 2 | 19324 | 9825 | 5512 | 2915 | 1515 | 727 | 364 | 209 | 110 | 60 | 35 | 15 |
| Replicate 3 | 19601 | 10382 | 5581 | 2870 | 1440 | 701 | 363 | 192 | 109 | 56 | 36 | 14 |
| Replicate 4 | 19104 | 10615 | 5662 | 2816 | 1464 | 707 | 401 | 185 | 97 | 55 | 39 | 14 |
| Arith. Mean | 1.92E+04 | 1.02E+04 | 5.50E+03 | 2.87E+03 | 1.49E+03 | 7.23E+02 | 3.85E+02 | 2.00E+02 | 1.09E+02 | 5.95E+01 | 3.70E+01 | 1.40E+01 |
| Arith. Std.Dev. | 3.62E+02 | 3.68E+02 | 1.80E+02 | 4.05E+01 | 4.73E+01 | 2.44E+01 | 2.49E+01 | 1.37E+01 | 8.66E+00 | 5.45E+00 | 1.83E+00 | 8.17E-01 |
| Arith. Net Mean | 1.92E+04 | 1.02E+04 | 5.49E+03 | 2.85E+03 | 1.48E+03 | 7.09E+02 | 3.71E+02 | 1.86E+02 | 9.45E+01 | 4.55E+01 | 2.30E+01 |
Stable 3. Fluorescein standard curve
| 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 | 0.297 | 0.227 | 0.126 | 0.231 | 0.186 | 0.141 | 0.152 | 0.221 | 0.117 |
| Colony 1, Replicate 2 | 0.282 | 0.213 | 0.102 | 0.167 | 0.147 | 0.111 | 0.109 | 0.182 | 0.066 |
| Colony 1, Replicate 3 | 0.261 | 0.222 | 0.104 | 0.164 | 0.146 | 0.112 | 0.103 | 0.206 | 0.041 |
| Colony 1, Replicate 4 | 0.254 | 0.061 | 0.102 | 0.158 | 0.145 | 0.105 | 0.104 | 0.178 | 0.036 |
| Colony 2, Replicate 1 | 0.278 | 0.232 | 0.113 | 0.176 | 0.15 | 0.107 | 0.109 | 0.201 | 0.035 |
| Colony 2, Replicate 2 | 0.299 | 0.221 | 0.105 | 0.178 | 0.151 | 0.113 | 0.107 | 0.221 | 0 |
| Colony 2, Replicate 3 | 0.256 | 0.216 | 0.121 | 0.157 | 0.151 | 0.111 | 0.107 | 0.199 | 0.037 |
| Colony 2, Replicate 4 | 0.267 | 0.217 | 0.118 | 0.161 | 0.149 | 0.109 | 0.11 | 0.212 | 0.041 |
| 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 | 0.48 | 0.471 | 0.495 | 0.663 | 0.542 | 0.52 | 0.321 | 0.566 | 0.041 |
| Colony 1, Replicate 2 | 0.491 | 0.481 | 0.493 | 0.644 | 0.543 | 0.532 | 0.331 | 0.574 | 0.046 |
| Colony 1, Replicate 3 | 0.458 | 0.438 | 0.48 | 0.662 | 0.534 | 0.538 | 0.332 | 0.567 | 0.054 |
| Colony 1, Replicate 4 | 0.523 | 0.535 | 0.536 | 0.701 | 0.588 | 0.563 | 0.361 | 0.599 | 0.064 |
| Colony 2, Replicate 1 | 0.497 | 0.495 | 0.478 | 0.65 | 0.561 | 0.551 | 0.355 | 0.61 | 0.039 |
| Colony 2, Replicate 2 | 0.512 | 0.486 | 0.512 | 0.665 | 0.563 | 0.549 | 0.354 | 0.612 | 0.037 |
| Colony 2, Replicate 3 | 0.484 | 0.451 | 0.497 | 0.681 | 0.563 | 0.56 | 0.337 | 0.591 | 0.035 |
| Colony 2, Replicate 4 | 0.478 | 0.501 | 0.49 | 0.676 | 0.569 | 0.552 | 0.347 | 0.201 | 0.036 |
Stable 4. Abs600 Raw Readings
