Professor John Paul
Lab 4 Entry
Field Trip to Pescadero State Beach
On September 19th, I arrived at the lab at 12:45PM. We then acquired a van and journeyed to Pescadero State Beach, south of San Francisco along the coast about 50 miles in hopes of seeing the yellow flowering Lupinus arboreus. Our professor then brought us towards the cliffside with sandy soil facing the ocean and showed us an example of Lupinus arboreus. These plants were scattered here and there along the cliffside but not in abundant numbers. There were no yellow flowers visible on this plant but it was still alive and well.
We then walked across the bridge and then under it in order to reach the other side of the road which was more inland. Here we also saw some Lupinus arboreus but in a little less sandy of soil. We were able to find some of the plants with flowers.
Other Lupinus arboreus were also located and we took pictures of them. They were about three to four feet wide and about two to four feet tall as mature plants.
After observing these Lupinus arboreus plants, we then headed back to the van and journeyed north towards San Francisco. We planned to also visit another beach ( I believe Pomponio State Beach) but due to time, we decided not to stop and look for more Lupinus arboreus because we risked getting caught in traffic. We arrived back at school around 4:45PM.
Professor John Paul
Lab 3 Entry
The Sushi Test (continued)
On September 12th, I returned to the lab and retrieved my PCR tubes that I put in the thermocycler on September 5th. Our group then brought our PCR tubes to the side benches in the lab and began setting up a new gel in order to do gel electrophoresis. One of my members (Kayla) used a micropipette and made four rows of three of 2 microliters of loading dye onto a small piece of parafilm. Kayla then added 3 microliters of each person’s PCR tubes in the correct corresponding rows of the droplets of loading dye, making sure to use a new pipette tip each time. I then used a microliter to transfer the 5 microliter samples from the parafilm into the wells of the gel, making sure to use different pipette tips each time and that the wells were labeled on a piece of separate paper and each person’s samples could be identified. Gel electrophoresis was then conducted for about 15 minutes.
Once approximately 15 minutes passed, our group collected the gel and took it to be observed under the machine for special imaging. The image showed that one of my group members had all three of their samples, two of my three samples worked (CP02 and CP03), and two of my members, unfortunately, had none of their samples work. My wells were the three on the far right on the top row.
Since two of my three samples worked (CP02 and CP03), I then continued with the ExoSap PCR Clean-Up protocol. I collaborated with a group of fellow classmates whose samples also worked. A master mix of containing water, 10x buffer, SAP, and Exo was made for 20 reactions and its combined total measured 250 microliters (12.5 microliters per reaction) and placed in ice. A long series of new PCR product tubes were provided by the professor and each of us received a certain number of these tubes (since I had two samples that worked, I got two tubes; my tubes were #10 for CP02 and #11 for CP03). I labeled and recorded my tubes with the appropriate identifications and proceded to transfer 7.5 microliters of my CP02 into #10 and 7.5 microliters of my CP03 into #11. Then I transferred 12.5 microliters of the ExoSap master mix into #10 and another 12.5 microliters into #11 (all while using clean pipette tips in between each transfer of liquids). Once each person finished establishing their PCR tubes, we all put our tubes together in a thermocycler. The lab period then ended and we were excused to leave. Our professor then put the tubes into a labeled tube rack and placed into the freezer.
On September 26th, we will continue with this lab and start the EXOSAP program.