Lab work for the week
This week in the lab has been the busiest week I have had here so far. On Tuesday I started the day off by making media and Nutrient Agar slants. These are composed of…
- Beef Extract
- Peptone
- Agar
- dH2O
After briefly cooking the agar in the microwave the media is pipetted out into test tubes, each containing around 10 mls of the Nutrient Agar. Everything is then placed into the autoclave which takes around 2 hours to run. I have also been tasked with conducting other tests on the endophytes we have cultured. These include: Salt resistance, motility, Catalase, nitrogen levels, and phosphorous levels. Before proceeding with the experiments I needed to make and prep all plates and broths for the project. Jensen’s Medium was made for the nitrogen detection experiment. Pikovkaya’s plates were made for the Phosphorus detection experiment but two of the ingredients were lacking so we are not too sure if it will work. NA slants for culturing were made, and Tryptic soy broth tubes were also made for the Motility test. Once everything was taken out of the autoclave we placed the slant tubes on a rack that allows a slant to form in the tube. Once they cooled and solidified we were able to streak some of the colonies from the plates onto these slants. My supervisor says that slants are typically used for working cultures, I have never streaked on slants before so it was exciting. We put them in the incubator and waited till the next day to check on them. For the motility test a colony from the endophyte strain was poked into the center of the tube about half the way down, we are then observing its growth. Wednesday was a holiday so there was no lab work being done, but on Thursday is when I started to make solutions for my slat resistance test. I made 4 strengths of it: 0 mM, 50 mM, 100 mM, 150 mM, and 200 mM of NaCl. Along with the NaCl solutions I also had to remake my Jensen’s Medium plates because they unfortunately didn’t solidify overnight which was due to using too much Magnesium sulfate 7H2O, and Ferrous sulfate 7H2O. So we remade the agar and put everything in the autoclave. While that was running I ended conducting the Catalase test which detects an enzyme in the endophytes that catalyzes the release of oxygen. In order to prepare this experiment I needed slides and 3% hydrogen peroxide. I then smeared a colony from the strains and placed a single drop of hydrogen peroxide on it and waited to see if it bubbles. If it bubbles then it has the enzyme that catalyzes oxygen, and no bubbles meant no enzyme. Out of the five strains I tested only two didn’t bubble, those included: In-b-590, and In-b-24. The fact that 24 didn’t bubble is due to the lack of a sample. In-b-24 is very hard to grow and I may not have a large enough sample to see. Finally Friday came around and I was able to conduct my salt resistance test! I only had 10 Nutrient Agar plates left so what I had to do is split the plates into 5 pieces and marked the different concentrations of salt. Then I mixed the colonies in 500 microliters of the salt concentration and pipetted 50 microliters onto the plate. I made two plates of each strain and popped those right in the incubator at 37 degrees C. Then since my Jensen’s Medium plates finally worked I was able to streaked on those and place them in the incubator over the weekend.
Weekend and Holiday activities!
As said above, Wednesday was a holiday so we didn’t have to go into work. So as a group we all headed to the town to the very small mall to pick up some stuff. I left my nice dress shoes at home so I ended up buying some $15 cute shoes for when we have to present them here! The others bought some clothes and a bag. After that Jessica N. has been telling us about this café that is super cute and super good so we went on a journey to find it. It was around a 10 minute walk from the main road, and we were pretty much walking through a cute little neighborhood. We finally stumbled upon and got ourselves into the cool AC building. The menu was great and I got a Spanish latte with cinnamon which was delicious. Once we were all done with our food and drinks we headed back. Typically we would have caught a bus back to IRRI but there were no more buses :(.
For the weekend on Saturday we decided that we will tackle the great Mount Makiling. The entirety of the hike starts at the base which you follow a manicured road up to the start of the hike to the peaks. But…. that would have added an extra 1 hour and a half to our journey so instead we paid 100 pesos to ride motorbikes up to the baes of the peaks! I have only ridden one motorbike before but this one was fun, although a bit scary especially since I felt us slip on a leaf. Once at the start of the trail there were some snack so I got a Turon for 10 pesos (it’s like banana lumpia but then coated in honey). Then we were off. I love hiking, it is the top most favorite of my hobbies. I hike a lot back home which is also a tropical rainforest environment, so this hike reminded me of home. There were many plants that seemed similar to those from home, for example they do have Lauhala (Pandanas) and there were so many ferns (if you know me ferns are my favorite). However there were also some plants that were super foreign, like trees which seemed to have some sort of parasitic fruit growing on them, or palms with thorns on them! There is also a large parasitic flower called Rafflesia. Sadly it wasn’t in bloom when we got to see it but they were still there. The one things that I did not appreciate on this hike were the leeches. We DO NOT have leeches on Kauai so when I got my first one of my thigh I freaked out. They were everywhere! Constantly biting, I was so happy that I have both long-sleeves and pants on. The hike was beautiful, maybe not the most spectacular scenery but the forest was stunning. If you sat quietly you could here the bugs humming and the bird singing, and if you listened close enough you could here some leaves clacking together similar to the olapa back at home. The guide who was with us would whistle a a very loud bird would respond, they seemed to be having a wonderful conversation! The dirt is similar to the mud in Pihea back at home, with clay in it which makes it more sticky and slippery. If it had rained this day we would have had an even harder time getting up. At the top of the peak which was peak two we could see the town but not much of it. The greenery is so thick and tall that we could only look at it through small clearings. Once we reached the rest point up top (station 30) there were butterflies dancing in the wind but then leeches crawling on the floor, I made the horrible decision to sit down up there and ended up getting a leech stuck to my butt (not fun at all). On the way down I also noticed how huge the tree ferns were! Overall the hike wasn’t super hard but I had a great time, it made me notice how much I miss home though. Once at the base of the mountain we got a jeepney back again and all took a much needed shower. I planned to sleep for the rest of the day, but no, because apparently two of the other interns wanted to come and visit Los Banos. I was tired, but still decided to go out. We ate at Seoul Kitchen where I got Sundubu jjigae which is a spicy tofu dish, it did have mussels in it so I gave it to Julia who has never had mussels! On Sunday our bodies were so sore that it ended up being a nice rest day. Hope you all have a wonderful week! A hui hou 🙂