InterLab
Preparation
We participated in answering the following question during the InterLab study of this year:
Can we reduce lab-to-lab variability in fluorescence measurements by normalizing to absolute cell count or colony-forming units (CFUs) instead of OD?
Materials
DNA/Plasmids
- Negative Control: BBa_R0040
- Positive Control: BBa_I20270
- Test Device 1: BBa_J364000
- Test Device 2: BBa_J364001
- Test Device 3: BBa_J364002
- Test Device 4: BBa_J364007
- Test Device 5: BBa_J364008
- Test Device 6: BBa_J364009
Apparatus
- 96 well plates (provided by Peking University)
- Plate reader
- Foil covered 50 ml tube
- Eppendorf tubes
- Pipettes
Materials
- LUDOX CL-X
- Silica beads
- Fluorescein
- Phosphate buffered saline
- LB media
- Chloramphenicol
- LB plates
- distilled water
Protocols
We followed the protocol provided by iGEM HQ so that inter-laboratory errors can be reduced. Protocols we used can be found here:
2018 InterLab Plate Reader ProtocolHelp: Protocols/Transformation
Results
I.Calibrations
OD600 reference point:
![OD<sub>600</sub> Reference Point](https://static.igem.org/mediawiki/2018/a/af/T--BJRS_China--OD600_Reference_Point.png)
Particle Standard Curve
![Particle Standard Curve](https://static.igem.org/mediawiki/2018/e/ef/T--BJRS_China--Particle_Standard_Curve.jpg)
Particle Standard Curve(log scale)
![Particle Standard Curve(log scale)](https://static.igem.org/mediawiki/2018/d/dd/T--BJRS_China--Particle_Standard_Curve%28log_scale%29.jpg)
Fluorescein Standard Curve
![Fluorescein Standard Curve](https://static.igem.org/mediawiki/2018/9/95/T--BJRS_China--Fluorescein_Standard_Curve.png)
Fluorescein Standard Curve(log scale)
![Fluorescein Standard Curve(log scale)](https://static.igem.org/mediawiki/2018/9/97/T--BJRS_China--Fluorescein_Standard_Curve%28log_scale%29.png)
II.Cell Measurements
Fluorescence Raw
![Fluorescence at 0h](https://static.igem.org/mediawiki/2018/0/06/T--BJRS_China--Fluorescence_Raw_0_hour.jpg)
![Fluorescence at 6h](https://static.igem.org/mediawiki/2018/1/1d/T--BJRS_China--Fluorescence_Raw_6_hour.jpg)
Abs600 Raw
![Abs600 at 0h](https://static.igem.org/mediawiki/2018/5/59/T--BJRS_China--Abs600_Raw_0_hour.jpg)
![Abs600 at 6h](https://static.igem.org/mediawiki/2018/8/86/T--BJRS_China--Abs600_Raw_6_hour.jpg)