Wetlab
EcnB part extracted from A(ABEF) synthesized plasmids and purified. It was then ligated with H backbone (pSB1C3) and transformed. Had some issues growing cultures with ligations. We are excited to start cloning!

Drylab
Dry lab discussed and finalized a design as well as the dimensions for the fish tag. Parts were designed through SolidWorks in member’s own time.

Policy and Practices
We began searching for fish farm contacts, and started our YOURS mentorship program. We introduced our mentors to the kids and taught them about DNA and its importance.

Wetlab
Internal (PstI) cut sites also found in AA & AE (switched to cutting the backbones and inserts with EcoRI and SpeI). AA+H (ZA) construct and AA+B (ZB) construct were sent for sequencing to verify the contents of the construct. The two primers added resulted in overlap, and sequencing was redone with one primer.
Flavo Subteam
Flavobacterium were re-plated, but they showed no growth after three days. Their growth is necessary to conduct ZOI (zone of inhibition) testing.

Drylab
We came up with and finalized the design of an applicator for the fish tag that improves upon the existing design. Parts were assembled through SolidWorks, and we planned to use the machine shop to build a metallic applicator.

Policy and Practices
We continued the topic of DNA in bacteria with the YOURS kids by talking about antibiotic resistance and how we only want certain genes in our own experiments. The kids learned how to pipet and load something into a gel and watched as we showed them how we run gel electrophoresis experiments everyday.

Wetlab
We had to re-culture several plasmids (pre-ligation). We also received and made glycerol stocks of five new plasmids.
Flavo Subteam
Flavobacterium growth was tested on different types of plates to see if our plates were the issue causing lack of growth.

Drylab
To market the product, the tag needs an appealing name. We brainstormed many potential ideas that may be useful in constructing a name. In addition, we researched specific tubing, clamps and other materials that we intend to utilize for the fish tag

Policy and Practices
We moved on to the topic of proteins to start to relate what we were introducing to YOURS kids with our own project so they would understand the relevance of our research. After discussing the basic properties of proteins, we helped them carry out an experiment that separated proteins from different dairy products. The visual results helped them better understand how chemical reactions can change molecular structures.
We contacted Bath hatchery and set up a meeting time in July so that we could drive there and talk to representatives in person and our project in perspective of current solutions.

Wetlab
ZE construct was sequenced and stored as glycerol stock along ZK, ZO, ZA, and ZB. The glycerol stocks are for later use in ZOI testing.
Flavo Subteam
Flavobacterium plates displayed lawn growth that will be used for ZOI tests.

Drylab
Dry lab continued with construction of the box. In addition, fish tags were being printed using high resolution 3D printers.

Policy and Practices
We took the YOURS kids to the Bathe hatchery. We spoke to Bob Sweet who explained how they keep an eye on various conditions in the fish's environment through the use of chemicals. He talked to us about the fish they raise, the types of diseases the fish face such as ferunculosis, and the overall day-to-day things the hatchery has to worry about like maintenance. The YOURS students also learned about how the fish are raised and transported in detail and were allowed to feed the fish. We learned that the hatchery has chosen not to use fish tags and instead clips fish fins to keep track. Sweet’s opinion of the future for the fish/agriculture industry was that the environment would put greater stresses on wild fish and therefore depending on farm-raised fish would be beneficial in supplying the demand for more food in a world with a rising population. We were also directed to speak to Andy Norse at Rome Hatchery who was more aware of chemicals used to treat water to prevent diseases, and we followed up with a call on Monday to plan a Skype meeting with Andy.
On Saturday, Grace and Saie went to the local Ithaca’s Farmer’s Market to find locals with opinions about GMOs and fish farming for our Humans and SynBio Facebook page.

Wetlab
We successfully sequenced and glycerol stocked ZR, ZL, and ZJ constructs. There were some weird results on gels of other constructs, possibly due to mutations in the backbone (H). We also started cloning MBP with Z constructs to stabilize the Ecn-B peptides since they were likely being degraded in the cell.
Flavo Subteam
Chloramphenicol ZOI control didn’t work. We attempted ZOI test with ampicillin instead for positive control data.

Drylab
Meeting addressed fundamental parts of a business plan. Members conducted research into various aspects of the aquaculture industry, consumer behavior, and finances in order to construct a comprehensive business plan.

Policy and Practices
iGem members Sachi and Saie presented at the The Summer Institute for Life Sciences (SILS) symposium. SILS is a program of the Office of Undergraduate Biology (OUB) at Cornell that invited undergraduates to present their summer research activities. These iGem members were able to talk about our research this summer with fishPharm.

Wetlab
We attempted Gibson Assembly with MBP-TEV insert and H (pSB1C3) backbone as an alternative cloning mechanism. The longer sequence will increase EcnB peptide stability.
We also received synthesized plasmids from DNA 2.0 with all EcnB inserts from biobricked Z constructs and MBP-TEV (not in pSB1C3).

Drylab
Dry lab continued with the design of labels and business cards. App development is proceeding without hurdles. The business plan was finalized.

Wetlab
We encountered issues with getting past ligation stage with synthesized plasmid inserts and H backbone.
To confirm EcnB expression, several SDS-PAGEs were done on constructs without stabilizing proteins to use for Western blots (to see their expression) and later ZOI assays.

Drylab
Business plan was reviewed by a professor with expertise in the area. Additional tags were printed, and samples were sent to prospective customers for feedback.

Policy and Practices
We sent our prototype of the fishBit to Rome Fish Hatchery and the Cornell Biological Field Station for feedback. We hope to use any advice and critiques to update our fishBit to fit the industry’s needs in the best way possible.

Wetlab
The second set of western blot revealed expression of ZG, MJ, and ZT. ZG was taken to be the positive control based on previous western blot. A series of ZOI disk diffusion assays were set up at the Cornell Vet School to test the efficacy of our BioBrick constructs.

Drylab
Dry lab tested the durability of fishBIT using agar gel samples and dead fish samples. In a hydraulics lab, fishBIT was tested for its resistance to current turbulence.

Policy and Practices
We sent our prototype of the fishBit to Bath Fish Hatchery. That same week, both hatcheries and the field station were able to give us valuable critiques including their opinion on the size of our product, the ease of application, and even encouraged us to continue with what they believed to be a valuable contribution to improvement of the fish industry.