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Latest revision as of 23:54, 17 October 2018

BJRS

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

  1. Negative Control: BBa_R0040
  2. Positive Control: BBa_I20270
  3. Test Device 1: BBa_J364000
  4. Test Device 2: BBa_J364001
  5. Test Device 3: BBa_J364002
  6. Test Device 4: BBa_J364007
  7. Test Device 5: BBa_J364008
  8. 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 Protocol
Help: Protocols/Transformation

Results

I.Calibrations

OD600 reference point:

OD<sub>600</sub> Reference Point
Table 1. OD600 Reference Point.

Particle Standard Curve

Particle Standard Curve
Figure 1. Particle Standard Curve.

Particle Standard Curve(log scale)

Particle Standard Curve(log scale)
Figure 1. Particle Standard Curve(log scale).

Fluorescein Standard Curve

Fluorescein Standard Curve
Figure 2. Fluorescein Standard Curve.

Fluorescein Standard Curve(log scale)

Fluorescein Standard Curve(log scale)
Figure 2. Fluorescein Standard Curve(log scale).

II.Cell Measurements

Fluorescence Raw

Fluorescence at 0h
Table 2. Raw Plate Reader Measurements of Fluorescence Raw at 0 hour.
Fluorescence at 6h
Table 3. Raw Plate Reader Measurements of Fluorescence Raw at 6 hours.

Abs600 Raw

Abs600 at 0h
Table 4. Raw Plate Reader Measurements of Abs600 Raw at 0 hour.
Abs600 at 6h
Table 5. Raw Plate Reader Measurements of Abs600 Raw at 6 hours.