Week 4 (2009)

This past week was particularly exciting because my frontline mentor, another graduate student, and I went to the Oregon Institute of Marine Biology (OIMB) for 2 days. OIMB is another research facility located on the coast near Coos Bay. There, we collected B.pacifica, another bryozoan that is closely related to B.neritina but is typically more difficult to work with. Also, judging from the number of publications online on B.pacifica, this bryozoan has been given less attention than B.neritina.

During our stay, we collected B.pacifica and preserved it in 4% paraformaldehyde and RNAlater for future analysis using FISH and DNA extractions/PCR, respectively. Also, we made an effort to collect B.pacifica larvae - these larvae are extremely small and very difficult to see! Bright lights stimulate B.pacifica to release their larvae into the water. So, we stimulated the larval release by keeping some B.pacifica we collected in the dark for 8+ hours, then we placed bright lights over the organisms. Sure enough, we began to see larvae swimming towards the light. They normally swim for up to 12 hours (according to the literature), until they finally settle on an appropriate surface and begin to develop. Anyway, we spent a few hours squinting at the water, armed with pipettes and small tubes to place the larvae in. Collecting them proved to be quite a challenge, but we managed to accumulate enough to do FISH on the samples and hopefully locate E.sertula!

Traveling to OIMB was also great because the research facility is very accomodating to other researchers. We were provided with a small cottage (it was so cute!) and a lot of lab space to work with. Also, the graduate student that was hosting us, Maya, was very helpful during our collection of the bryozoans. Overall, it was a very pleasant experience! 

For the remainder of the week, I extracted DNA from B.pacifica and used PCR to amplify the 16S rRNA gene. We tried a variety of different primers, from the general bacterial (1492r and 27f) to more specific E.sertula primers. From my first two PCRs, it appeared that the DNA I extracted is a bit degraded. So, at the moment, I have a third PCR in progress using a combination of new primers that will hopefully amplify a smaller portion of E.sertula specific genes (I'm hoping that the DNA isn't so degraded that this small portion won't amplify! We'll see...)

Finally, I wanted to mention the NSF site visit that took place this week. From watching all the presentations, it seems that CMOP is doing a really good job of using its funding wisely; the facility is also tackling very appropriate and difficult problems that are relevant to this Pacific Northwest. Congrats CMOP for another year of good work!

GOALS FOR NEXT WEEK: 
- Do FISH on the B.pacifica larvae
- Do FISH on the B.pacifica ovicells
- Take a closer look at the FISH results from the B.neritina ovicells with my frontline mentor