Junior Year

Journal 6: (9/30/26)

While I’m waiting for my yeast plates to come, I’ve been working on the Soracle. The Soracle will be a network of life science sensors (air quality, temperature, etc.) based on ESP32s and coded by Arduino systems. Here is a link to a doc I’m using to plan and keep the names of specific sensors.https://docs.google.com/document/d/1AMZCXtzkdM-4fzOn2vF-w1YS1zMZPOAZJHwkdcfUjiw/edit?tab=t.0

The ESP32’s sensors have wifi and Bluetooth and will be able to do constant updates. They can broadcast to a section of the WISRD website where we can compile all life science project data.

Journal 5: (9/16/26)

I have been practicing face painting in WISRD for the last few class periods in preparation for Dark Matter Day. I will run the face-painting booth again :)

Journal 4: (9/15/26)

I found a website that explains the functions of specific yeast genes!! For size specifically, I can wait until my experiment comes back and the bigger yeast cells survive. Then do some guess-and-check with genes and the PCR machine to see which gene is making the yeast “immune” to the microplastics. (I know the bigger yeast will survive because of Parker’s experiment.)

https://www.yeastgenome.org/phenotype/increased_cell_size

Another sequence website:

https://www.uniprot.org/citations/7813418

Journal 3: (9/14/26)

Ok, Megan helped me write the protocol I will be using from Parker’s poster. I had no clue how to decipher it, so thank you Megan 😭. I emailed Megan asking for the materials I need, and it will probably take a few weeks to arrive. Almost everything I need we have IN THE FRIDGE. REMEMBER THAT. Also, I found a CRISPR Program!:

https://crispor.gi.ucsc.edu/

And here is a website for ordering gRNA:

https://www.synthego.com/products/crispr-kits/synthetic-sgrna/?utm_term=guide%20rna%20design&utm_campaign=sgRNA+Products+Campaign&utm_source=adwords&utm_medium=ppc&hsa_tgt=kwd-814961122226&hsa_grp=176530615571&hsa_src=g&hsa_net=adwords&hsa_mt=p&hsa_ver=3&hsa_ad=742172437665&hsa_acc=6964378581&hsa_kw=guide%20rna%20design&hsa_cam=11155205430&gad_source=1&gad_campaignid=11155205430&gbraid=0AAAAADcUr1TfIvT-ZC7BTdSHEsrggUHx-&gclid=EAIaIQobChMIxau9qMrulgMVfAe8AR1DihEHEAAYASAAEgJ1c_D_BwE#grnadesign

Here is the protocol:

  1. First, turn on the incubator so it is heating to 35°C, and sterilize the laminar flow hood with 70% ethanol

  2. In the flow hood: 

    a. Rehydrating the yeast: Sprinkle dried Saccharomyces cerevisiae Type 2 (Baker’s yeast) onto the surface of a sterile Petri dish filled with water heated in a kettle to 95°F and let it sit for 15 minutes, then stir slightly and leave for another 5 minutes.

    b. 1 mL of the rehydrated yeast was then micropipetted using a wide-bore pipet tip (created by snipping the tip off a 1000 ul tip) onto 2 Yeast extract dextrose (YED) agar plates. Do this for as many plates as you will need, plus a couple extra just in case.

    c. Let the YED plates sit for a minute so the yeast can adhere to the agar, then place them in an incubator upside down at 35°C for 18-19 hours; these plates are the base plates. 

    d. After the incubation time has passed, a thin layer of yeast colonies should be present on the YED plates; check the plates for obvious contamination, and if it is seen, dispose of the plate.

  3. Before exposing the yeast to microplastics, we get a base density of cells. 

    a. Again in the flow hood, use a sterile swab to lightly scrape some yeast cells off the base plate. Swirl the swab in a well of a spectrophotometer plate that is filled with sterile water, and fill the well to the brim with more sterile water. The plate was then put into the spectrophotometer at 400 - 700 wavelength, and the reference data was recorded to compare against after the experiment. 

    b. Use a wide-bore micropipette to transfer the yeast solution from the two wells into two centrifuge tubes, keeping track of which is which. Gently shake the tubes to make sure the yeast is distributed evenly through the tube. 

    c. Exactly half of the solution is then pipetted onto a YED plate, and these plates are then placed agar-side down in an incubator for 24 hours to establish the yeast on the plate.

  4. After the 24-hour incubation period, add  20μl of the solution of latex beads is then pipetted onto the center of one of the plates. Then both plates were placed upside down (agar side on top) for 6 days in the incubator

  5. After this six-day incubation period, the plates are taken from the incubator, and a sterile swab is used to scrape some of the yeast into a fresh spectrophotometer plate with sterile water once again.

    10 µL of a propidium iodide solution is added in both wells, and the plate is measured in the spectrophotometer. This pattern is then repeated, with samples being taken from each plate, measured, and placed into a new plate to grow, going back to step h.

And here is the stuff I need:

Around 10-12 YED Plates (yeast extract dextrose)

Spectrophotometer plate? Which we might already have?

Journal 2: (9/1/26)

I am learning how to use our PCR machine and how PCR works in general. What I’ve figured out is that PCR testing is basically guess-and-check for certain genes. For example, I would take a section of yeast genes, and I insert a primer for a gene I suspect the yeast has; if that gene is produced in the PCR machine, I know that I was correct and the yeast contains that gene. If that gene is not produced, I was wrong and that gene does not exist in the yeast. I am also still struggling to contact Parker (SHE WON’T TEXT ME BACK I’M LOSING IT I NEED HER NOTES). But thankfully, Megan sent me Parker’s poster, and I have been trying to work backwards from her procedure to get a protocol. Unfortunately, her procedure is not very specific, so I am having trouble getting an exact list of steps. I’m confident I will be able to decipher it, though, especially with Megan’s help. I also really hope Parker texts me back.

Journal 1: (8/24/26)

It’s the start of WISRD! I am working on finding Parker’s protocol for the Yeast Genetic Engineering Lab; as soon as I do, I’m gonna jump in. So far, I have been reorganizing my journal and getting re-acclimatized to WISRD. I am planning to pitch some plans to make WISRD more interconnected at next Thursday's WISRD meeting.