The Interlab study of Team Hong_Kong_JSS is a collaborative effort together with Team Hong_Kong_CUHK during 17 to 23 of July. we are extremely grateful that Team CUHK provided us with material and equipment assistance, as a high school team like us lacks access to equipment necessary for this study, such as the plate reader. We only participated in the Plate reader stream.

Materials

Plate reader: BMG labtech CLARIOstar

96-spot well plates: clear plates from Corning

Bacteria strains: E.coli DH5α

Test devices: obtained from the distribution kit

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

#The 50ml falcon tube used in the experiments are clear tubes, but they are covered under an amber cover from the shaker.

Methods

The protocol we followed is given by the iGEM headquarter for all interlab-participating teams to follow.

Here’s the link https://static.igem.org/mediawiki/2018/0/09/2018_InterLab_Plate_Reader_Protocol.pdf

Results-Calibration

Calibration 1 OD600 Reference point

Fig 1

Calibration 2 Particle Standard Curve

Fig 2

Discussion

We noticed the general increasing trend of absorbance for the particle curve, but the data does not behave as a perfect straight line. We noticed that the silica beads condensed and clustered together very quickly upon the mixing of the solution (by pipette up and down). Therefore, we keep the 96 well plate on a vortexer to keep shaking it gently until putting it in the plate reader. However it seems like the silica still clustered at the bottom and this might make the readings inconsistent and causes some error.

Calibration 3 Fluorescence standard curve

Fig 3

Results-Cell measurement

Raw plate reader values

Fluorescence measurement

Fig 4

Absorbance Measurement

Fig 5

Net measurement values

uM Fluorescein / OD

Fig 6

Net Fluorescein a.u.

Fig 7

Net Abs600

Fig 8

Results-Colony Forming Unit

OD 600 measurement of the 2 device

Fig 9

Colony Count

Fig 10

CFU Calculations

Fig 11

Acknowledgement

Participating in the interlab study was definitely a challenge to our team as we lack the necessary equipment (eg. the Plate reader) to carry out the measurement, and we would would like to thank team Hong Kong CUHK once again. Without their generous help, we would not have been able to participate in this study.

Conclusion

The interlab study was definitely quite challenging for a high school team like us, as we lack the equipment and experience in running a experiment in this size. Some of the apparatus and reagent (eg. the Amber 50ml Falcon tube) prescribed in the protocol were not available in both our lab and CUHK’s lab, so we are often to adapt and use various alternative methods. However, this has been an amazing experience for us to improve our wet lab work ability and we are so grateful to be able to participate in this research.