Working in the Lab!
We once again had a long weekend with Monday being a holiday so my work week didn’t start until Tuesday. My supervisor had some errands to run so he was off for the entire day which meant no wet lab activities for me. However my day was full of researching literature instead! I was tasked with finding methods on extracting and measuring the components of exudates from roots. There was a lot of reading to be done but by the end of the day I had methods for bonds extraction and analysis. The methods to extract from soil typically may involve some suction and cultures of that soil in a nutrient broth. For roots you soak the root for 48 hours and then switch out the culture and soak for another 48 h. Exudates should be in the media after the 48 hours, all one has to do then is run the solution through a filtrate to only be collecting those exudates. Within the exudate extract there is a mixture of acids, bases and neutrals, in order to divided them one must use chromatography and columns. After my readings were complete the last task for the day was to water the rice seedlings for our experiment in the greenhouse. We won’t be using these plants as the main samples for our project they are for backup purposes only. on Wednesday is when we really started to do more things. Once my supervisor came into lab we made some media for our project. Typically there is a lab meeting on Wednesdays at 1pm but this week it was pushed back until Thursday. Right after lunch I was instructed to make 500 mls of Nutrient agar which consists of peptone, Beef extract, and agar. When that was completed I also had to make Nutrient broth which is the same as nutrient agar just without any agar added to the broth. once those were mixed up we placed them in the autoclave which typically runs for 2 hours. it was now around 2 pm by that time. While the autoclave was running, my supervisor, also the head of this project, took me to the greenhouse to transplant our seedlings of rice into bigger pots for growth. It was nice to get out of the lab, but those greenhouses are so hot. When I went to grab the last bit of plants I ended splattering mud up my shirt, it wasn’t too much and came off right when I washed it. Once we finished up our gardening for the week, we headed back to lab to see if the autoclave was complete. Unfortunately is was not which meant I would have to wait until tomorrow to pour my nutrient agar plates. Once Thursday hit we started right at 8am to get that agar from yesterday melted using a hot plate. the melting took around 30 minutes but then it was time to pour. Once I poured we realized that the agar wasn’t cooled down enough so it made some precipitation on the plates meaning i couldn’t streak endophytes until Friday. The reason we had to make Nutrient Agar plates is because four of the five endophytes that we chose from the freezer to be revived can only grow on Nutrient agar and not Kings B Medium. While we waited for the plates to cool we got in-b-24 endophytes out of the freezer and streaked it. These endophytes are stored in skim milk and glycerol so they had a sweet smell every time I opened the tube. once placing the plates in the incubator it was time to head to the lab meeting. I was instructed to crate a presentation about what I will be working on here in lab during my stay however, there were two other presenters and we didn’t finish the meeting until 3pm. Since my nutrient Agar plates were still drying we decided that we will streak the rest of the endophytes tomorrow morning. On Friday we started wet lab activities once again right at 8am. First we had to defrost the endophytes from the freezer which meant to leave them on ice for about 20-30 minutes. During that time I helped IA (my supervisor) with some more media he was preparing. Once the skim milk and endophytes were melted I vortexed them juts a bit to make sure it was mixed through all the was to ensure that I will be picking up endophytes and not just sugars. I streaked two plates per endophyte samples which cam out to a total of 8 plates. Once those were done it was finally lunch time. after lunch my supervisor had to run some errands again and I was told to find other biochemical tests I would like to conduct on the endophytes. Some that we are already doing are Nitrogen and Phosphorous levels. I read some articles and other test include Potassium levels, possibly drought and salinity tests and many more. I still have to run them by my supervisor to see exactly what we will be testing. I look forward to isolating more endophytes next week.
Weekend Activites!
After completing all my work I was able to get off of work an hour early which was helpful because the we were going to Manila for the weekend and our bus was picking us up at 5:30 pm. We finally got to Manila around 8pm and checked into our hostel. I have only stayed in one hostel before and this one was quite nice compared to the other one. We booked a 6 person room which meant that we were the only people in that room. The next day we got up around 9 and didn’t head out until 11am, we decided to head down to the café right below our hostel which was called Sasa Café. it was cute and the food and drinks were delicious! After wards we headed out to explore some of the museums. Sadly however the only museum we got to explore was the anthropology museum, I noticed that the group with us (Jessica, Julia, Jessica) very much enjoy Museums and wanted to make sure we saw everything. After about many hours in that Museum we wanted to head to the Mall Of Asia Also known as MOA to meet up with some of the others from FNRI. We spent many more hours in the mall where I bought some skin care products and of course food. After our shopping we were all so hungry so we had dinner at a cute ramen shop whcih had some very delicious ramen. Later we headed back to our hostel. Sunday was our last day in manila and the grils went to church but I am not one who goes to church often so instead i went to the mall to get some food with Sam. after spending a little while there we headed back to our hostel to check out and get picked up by our bus to head back to IRRI. And that was the weekend!
