Slow week in S-STEM

This week in S-STEM, I don't have much to report. Last week I finished my agar plates and this week I have been waiting patiently for my algae to grow. I expected to see little colonies forming, but all I see is a faint haze. I am a little concerned that they won't grow, so I'm crossing my fingers.

Tomorrow, we're taking a field trip to the Wildlife World Zoo so I'm pretty excited for that. I went recently with my dad and my daughter, and ever since she has been begging me to go. Luckily for her, she'll be coming along to see the "mingos" aka flamingos, her favorite. Plenty of pics to come!

Relly and grandpa at Wildlife World Zoo

Agar plates and Algae die-off cont..

This week in STEM I was finally able to prepare my plates, but now both of my algae strains are dying off! I have already refreshed the media for Arthrospira but it has yet to have a big bloom like when I started my culture. Now it is time to refresh my Nannochloropsis media because it is starting to turn brown as well. I hope to be able to plate both strains tomorrow to allow them to grow over the long weekend. I was concerned that I wouldn't be able to get my agar to set right because of my issues last week. I fully melted the agar before refrigerating the culture media, but the strange grains persisted. I cooked my agar until it foamed up so it may be because of the salinity. Luckily, when I checked my agar media before autoclaving it appeared normal and unaffected from the grits I couldn't dissolve. Autoclaving did not fully melt my agar, I had a strange bubbly lump inside my flask so I put it back into the autoclave on the "melt" setting.  Hopefully this will work!

Solid agar media

Dying algae culture

Algae Die-off and Chunky agar

Last week at STEM I started work on my agar plates but I found that some of my algae had stared to die off. After panicking for a moment, I remembered that I could refresh my culture with fresh nutrient-fortified solution. The Nannochloropsis culture looked great, but my Arthrospira water started looking brown. I also prepared salinated water for my isolation agar plates. I added 31.25g of Instant Ocean to 1L of DI water with 1 vial of Alga-Grow concentrate to make the liquid portion for my plates with 10.00g of plant tissue culture grade agar. On the package for Instant Ocean the recipe was for a gallon, so I converted it into g/L, but in hindsight, I should have just made the gallon and poured out a liter. As it heated, there appeared to be chunks of agar that wouldn't melt but I autoclaved it anyways. When it was done, the chunks still persisted so I tried to melt it on the hot plate/stirrer and subsequently burned it. That was pretty disappointing, so I prepared another liter of agar culture and put it into the fridge for this week.

Spirulina dying

Chunky agar

Algal Research Project

For my research project this semester, I wanted to do something regarding algae with a focus on genetics. So, I came up with a toxicity bioassay based on J.I. Nirmal Kumar's study, "Toxicity analysis of pesticides on cyanobacterial species by 16s rDNA molecular characterization."

Once I isolate single colonies of my algal species, Nannochloropsis sp. and Arthrospira platensis, I will do a PCR (polymerase chain reaction) of the 16s RNA gene. Then I will subject the cultures to increasing amount of foliar pesticide, tebuconazole, similar to Kumar's study. However, I will be using a home use formula of the same brand, Kumar used a commercial agriculture formulation. The idea is to continue doing a PCR and record any mutations in the RNA structure; such as insertions, deletions or mismatching of base pairs.

Here are a few photos of the starter cultures.

citation: Kumar, J.I. Nirmal, et al. "Toxicity analysis of pesticides on cyanobacterial species by 16s rDNA molecular characterization." Proceedings of the International Academy of Ecology and Environmental Sciences 3.2 (2013)

Glad to be Back

Hi everybody! It feels really good to be back and I hope to hit the ground running, so to speak. I have decided to do my project on the toxicity of pesticides on cyanobacterial species; in this case Nannochloropsis and Arthrospira aka Spirulina. Matt has already ordered the cultures, so hopefully things go smoothly while I try to grow my cyanobacteria. Since it may not work out, I may do a back up project concerning algae. I'm not sure what at this point, so cross your fingers for me! I look forward to seeing you all at orientation Friday. Here's to a great semester! :)

Photo: http://www.ucmp.berkeley.edu/bacteria/spirulina.jpg