Summary of Monthly Goals:
- Collect more tunicates and maintain them in recirculating seawater system
- Continue troubleshooting through primary cell culture protocol
- Decide on a chemical mutagen to use for colony exposures
- Get an experimental design draft completed for the in vivo colony exposures
Collecting more Botryllus schlosseri
Before leaving for the winter break, the Gardell Lab reduced the number of colonies kept in the recirculating seawater system. After checking on them today, none of the colonies kept over the break are alive. As such, before getting started on colony exposures or working through cell culturing, we will need to re-establish a colony stock in house. I plan to collect more colonies from the De Moines Marina. I want to incorporate dipping colonies in Coral Rx upon arrival to UW Tacoma to improve survival.
Ideally the colonies used for exposures and cell culturing are lab grown but it is still not know if colonies are not proliferative in our lab conditions due to the season or some other constraint.
Troubleshooting primary cell culture protocol
Prior to the end of Fall Quarter I began working through the primary cell culture protocol Valerie (UCD) and I drafted. However, due to some misinterpretation, I was not able to complete the protocol. I plan to work through the protocol again with a fresh batch of colonies.
Chemical mutagen for colony exposures
I have been reading through the literature to learn more about genotoxic chemicals that may be of interest for both the colony exposures and cell exposures. A couple of mutagens we potentially are interested in are benzene[a]pyrene and nickle chloride. Particlarly interested in chemicals capable of clastogenic damage.