Difference between revisions of "Team:Reading/Experiments"
| Line 5: | Line 5: | ||
<p> This section documents the designs and protocols for experiments we carried out during our project </p> | <p> This section documents the designs and protocols for experiments we carried out during our project </p> | ||
<p><h5>Competition assay</h5></p> | <p><h5>Competition assay</h5></p> | ||
| − | <p> | + | <p>A competition is a simple experiment to provide information on the safety of the GM cyanobacteria we propose to use in our fuel cell.</p> |
| + | <p>The design of the experiment is as follows, observe aseptic technique throughout, 6803 refers to Synechocystis sp. PCC 6803: | ||
| + | <ul> | ||
| + | <li>Add approx. 200ml of BG-11 medium to a sterile conical flask.</li> | ||
| + | <li>Inoculate flask with 10µl of stock Wild type 6803, and 10µl of stock mutant 6803.</li> | ||
| + | <li>Incubate flask in incubator on a shaker, at 58rpm and 30°C, under bright white light. Allow bacteria time to grow, monitoring the growth over time by observing the colour of the medium and by taking OD750 readings.</li> | ||
| + | <li>Once the flask had reached a suitable cell density (e: g hundreds of millions of cells per ml), spread 1ml of culture over the surface of two plates, one of BG-11 and agar, and one which additionally contains the antibiotic Kanamycin (mutants should be marked with a Kanamycin resistance gene).</li> | ||
| + | <li>Estimate the colony forming units per ml (cfu/ml) on each plate.</li> | ||
| + | <li>The difference in cfu/ml on each plate will give an indication of which strain of 6803, wild type or mutant, will out-compete the other when placed in the same environment.</li> | ||
| + | </ul> | ||
| + | </p> | ||
<p><h5>Ferricyanide assay to measure electron output of 6803</h5></p> | <p><h5>Ferricyanide assay to measure electron output of 6803</h5></p> | ||
<p><h5>Public perception and awareness study: Methods and Questionaire</h5></p> | <p><h5>Public perception and awareness study: Methods and Questionaire</h5></p> | ||
Revision as of 17:20, 18 August 2015
Experiments
This section documents the designs and protocols for experiments we carried out during our project
Competition assay
A competition is a simple experiment to provide information on the safety of the GM cyanobacteria we propose to use in our fuel cell.
The design of the experiment is as follows, observe aseptic technique throughout, 6803 refers to Synechocystis sp. PCC 6803:
- Add approx. 200ml of BG-11 medium to a sterile conical flask.
- Inoculate flask with 10µl of stock Wild type 6803, and 10µl of stock mutant 6803.
- Incubate flask in incubator on a shaker, at 58rpm and 30°C, under bright white light. Allow bacteria time to grow, monitoring the growth over time by observing the colour of the medium and by taking OD750 readings.
- Once the flask had reached a suitable cell density (e: g hundreds of millions of cells per ml), spread 1ml of culture over the surface of two plates, one of BG-11 and agar, and one which additionally contains the antibiotic Kanamycin (mutants should be marked with a Kanamycin resistance gene).
- Estimate the colony forming units per ml (cfu/ml) on each plate.
- The difference in cfu/ml on each plate will give an indication of which strain of 6803, wild type or mutant, will out-compete the other when placed in the same environment.