Team:iTesla-SoundBio/InterLab
Summary: For the 2018 InterLab study, high school students at iTesla-SoundBio prepared various colonies and dilutions to test for trends absorbance, fluorescence, and concentration. For controls we took the absorbance of silica beads that are similar to cell absorbance. Then we took the absorbance of fluorescein serial dilutions to get a trend line of different absorbances as concentration of fluorescence decreases.
Massive thanks to the Washington iGEM InterLab team for guiding us through the protocol and lending us their plate readers.
Below, is the timeline that we followed in completing the InterLab protocol.
Part One- LUDOX Calibration Protocol
96 Well Plate Table
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | |
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| A | LUDOX | ddH2O | ||||||||||
| B | LUDOX | ddH2O | ||||||||||
| C | LUDOX | ddH2O | ||||||||||
| D | LUDOX | ddH2O | ||||||||||
| E | ||||||||||||
| F | ||||||||||||
| G | ||||||||||||
| H |
Here are the results for the Absorbance 600 of the LUDOX protcol
OD600 Reference Point Data
Part Two- Microsphere Protocol
Part Three- Fluorescein Protocol
Our experience with Interlab in one word would be rough. Our lab did not have a -80 degree freezer at that time so we had to store our competent cells at -20 degrees. One issue we had was when the InterLab leads were not present at the wetlab session to transform the E. coli cells with the devices, the members that were present used all the DNA that was provided. So when we had to re-do our InterLab protocols, we had to think of work arounds for the protocols. Additionally, we had to use the plate reader at Washington iGEM lab meaning that we had to schedule the wetlab days taking into account the waiting steps according to the schedule of the Washington iGEM team. Some possible sources of error or limitations were that we did not have black 96 well plates with a clear bottom. Instead, we used a 96 well plate that was clear throughout.