Dry Lab

• An overview of photosystem 2, the chlorophyll biosynthesis pathway, and the hydrogen production was given by our advisors. The areas which needed work were discussed.
• We started planning for creation of combined plasmid for hydrogen production by combining; Ferredoxin/Ferredoxin Reductase with the Hyd1 biobricks.
• Discussion on human outreach was started.
• Began talking to the SDU iGEM team on collaboration
• Discussed the creation of an iGEM Macquarie game
• Started planning for attending synthetic biology conference and ACUR (Adelaide).

Dry Lab

• India suggested the idea of a creating a game as her brother could help us out with that.
• We got in contact with Joanne Jamie, an academic at Macquarie University, to gain more information about National Science Week and the university’s Orientation Day for our outreach possibilities.
• She invited us to help out in the Chifley School Program (19th September) which we agreed would be better than the other two. We have a 1 hour time slot in which we can conduct wet and dry lab activities with gifted and talented high school science students.
• The team was approached by Nebraska to fill out their survey

Wet Lab

1. Make up new competent cells.
2. Run a gel of PCR products.
3. Transform cells using only 2ul HydEFG and new competent cells. Plate up these transormants.
4. Make overnight liquid cultures of successful HydEFG cultures.
5. Miniprep HydEFG from the above.
6. Digest and run of gel the resultant extracted plasmid to check if worth sending for sequencing.
7. If result from FER/Hyd A and B sequencing is unsuccessful then a new ligation of Fer/Hyd should be attempted. Look at what might need tweaking.

• Instead of making up new competent cells, we used commercial competent cells which proved successful as we were able to grow HydEFG transformants. Plasmids from these were extracted via miniprep. They were digested and run on a gel. The gel results were not as expected _____. Because of this, new colonies should be screened.
• The sequencing came back for Fer/Hyd A and B. Both were as confirmed. The glycerol stocks were then used to prep overnight cultures which were induced and tested for their ability to produce hydrogen. The Clark electrode was used to measure the H2 production and recordings went off the scale. This was great news. We are looking into the best ways to present this data.
• Unfortunately we still have not had any success with PCR amplification of backbones.
• HydEFG were sent to be sequenced and other cultures were grown as well as their glycerol stock.