Difference between revisions of "Team:Newcastle/Notebook/Lab/Lightbulb/Week 15"
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<h2>September 27th 2016</h2> | <h2>September 27th 2016</h2> | ||
| − | <p> | + | <p>Due to the issues with the current plate reader we were forced to change machines and were now using a BMG FLUOstar omega plate reader the same machine that was used in the inter-lab study. We continued to use the same plate layout as before. As we were using different plate reader, the gain values previously used had to be altered. Due to the higher fluorescence values compared to that with the inter-lab study, we set the gain value to 250. The plate was then run using a wavelength of 485nm excitation, 520nm emission for GFP and 600nm absorbance at 42°C. Inoculated fresh LB culture for use on the following day. </p> |
<h2>September 28th 2016 </h2> | <h2>September 28th 2016 </h2> | ||
| − | <p>We collected the data from the plate reader that was set up the day before and this time the samples had not saturated the plate reader although the signal was fairly noisy. Nonetheless we decided to continue with this protocol. The plate reader was again set up and this time ran at | + | <p>We collected the data from the plate reader that was set up the day before and this time the samples had not saturated the plate reader although the signal was fairly noisy. Nonetheless we decided to continue with this protocol. The plate reader was again set up and this time ran at 37°C. Inoculated fresh LB culture for use on the following day. </p> |
<h2>September 29th 2016 </h2> | <h2>September 29th 2016 </h2> | ||
| − | <p>Again the data we collected was noisy however, we decided to continue with the experiment and run the plate reader again but this time at | + | <p>Again the data we collected was noisy however, we decided to continue with the experiment and run the plate reader again but this time at 30°C. Inoculated fresh LB culture for use on the following day. </p> |
<h2>September 30th </h2> | <h2>September 30th </h2> | ||
| − | <p>We collected the final data set from the plate reader, which was again slightly noisy due to the low gain value of only 250. You can see our final results here. We plan to carry out the experiment once more at each temperature with a gain value of 1000.</p> | + | <p>We collected the final data set from the plate reader, which was again slightly noisy due to the low gain value of only 250. You can see our final results <a href="https://2016.igem.org/Team:Newcastle/Proof/ElectricallyInducedLightBulb">here.</a> We plan to carry out the experiment once more at each temperature with a gain value of 1000.</p> |
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Latest revision as of 20:52, 19 October 2016
September 26th 2016
Today we entered the lab full of anticipation to see the results from the plate reader set up the previous week. Unfortunately, the results were not as we hoped. Many of the wells had AGAIN saturated the detection limits of the plate reader. After talking to our advisers, we decided a more appropriate negative control was required so as a result from this point forward we used BL21 wild type E.coli cells; the new plate layout is as follows. We inoculated fresh liquid LB broth for use with the plate reader and testing the battery constructs:
- Battery 1
- Battery 2
- DNA2.0 light construct
- Alternative light construct
- Variable Resistor (tagged)
- Variable Resistor (un-tagged)
- Positive Control (from InterLab study)
- Sample 1 (from InterLab study)
- BL21 negative control
September 27th 2016
Due to the issues with the current plate reader we were forced to change machines and were now using a BMG FLUOstar omega plate reader the same machine that was used in the inter-lab study. We continued to use the same plate layout as before. As we were using different plate reader, the gain values previously used had to be altered. Due to the higher fluorescence values compared to that with the inter-lab study, we set the gain value to 250. The plate was then run using a wavelength of 485nm excitation, 520nm emission for GFP and 600nm absorbance at 42°C. Inoculated fresh LB culture for use on the following day.
September 28th 2016
We collected the data from the plate reader that was set up the day before and this time the samples had not saturated the plate reader although the signal was fairly noisy. Nonetheless we decided to continue with this protocol. The plate reader was again set up and this time ran at 37°C. Inoculated fresh LB culture for use on the following day.
September 29th 2016
Again the data we collected was noisy however, we decided to continue with the experiment and run the plate reader again but this time at 30°C. Inoculated fresh LB culture for use on the following day.
September 30th
We collected the final data set from the plate reader, which was again slightly noisy due to the low gain value of only 250. You can see our final results here. We plan to carry out the experiment once more at each temperature with a gain value of 1000.
