Advanced Math/Science Research (AMSR) is a yearlong course that offers students an authentic independent laboratory experience to design and execute an original research project of their design in the biological, physical, or social sciences. What sets Berkshire’s course apart is the program’s design: students interact with a professional scientist to conduct real-world research in world-class facilities. The course culminates with a research paper. STUDENTS often enter national science competitions and PRESENT THE RESULTS TO MEMBERS OF THE MATH AND SCIENCE DEPARTMENTS AND THE BERKSHIRE COMMUNITY.
STUDENT PROJECT 1: Melatonin for worms?
MOYA '21 (LEARNING REMOTELY from Thailand THIS YEAR )
With an interest in sleep cycles, Moya designed an experiment to test the effects of melatonin on the hibernative (sleep) state of the model organism C. elegans. She developed the hypothesis, designed the methods, we conducted the experiments on campus and sent her the data for analysis.
Planning out the experiment
- Independent variable: Amount of melatonin as nutrients (0-100mg/L of melatonin)
- Dependent variable: Time for C. elegans to reach dauer stage
Hypothesis: the more melatonin in the solution, the faster the c. elegans will leave dauer stage (the faster the beads will appear in the c elegans)
- Prepare the Nematode Growth Medium (NGM), Autoclave the solution
- Then add cholesterol, salts, and melatonin after the solution has been autoclaved
- Melatonin solution was made fresh (50 mg/ml) (500 mg in 10ml of EtOH) then filter-sterilized.
- Melatonin was spread on plates using the hockey stick method. Let absorb at room temp then moved to 4C
- Using a worm picker, place individual adult worms into a 5 separate petri dish with different dosages of melatonin (0, 25, 50, 75 and 100mg/L) and equal amounts of Nematode Growth Medium (NGM)
- Wait for the C. elegans to reach dauer stage (check daily)
- Add 0.1uL of fluorescent latex beads with the 100ul OP50 food and place them in the middle of the dish.
- Check daily and to see if the red beads are present by fluorescence microscopy (may need to pick worms and check if plates are too thick). **If they are not present then the worms are still in dauer stage but if the beads are present then they have gotten out of dauer stage
- Compare the data to see if more melatonin increases the time it takes for the C. elegans to develop.
Moya's study suggests that her hypothesis was incorrect and, in fact, the maturation from egg to mature worm is actually delayed by melatonin in a dose-dependent manner. Sample images are shown.
In progress: next steps are to use fluorescent beads described to distinguish between adult and immature worms and confirm light microscopy
STUDENT PROJECT 2: stress is good (for viruses...)
Ava '21 (learning in person this year)
Ava is a Two year AMSR Program Student and PARTICIPATED IN 4 YEARS OF THE AFTERNOON AMSR PROGRAM. With an interest in virology, Ava wanted to follow up on some unpublished data suggesting that some viruses need stress to execute their gene expression cycle.
Ava has been studying stress-activated transcription factors
click below to Listen to AVA describe the method she is using to study Nrf2
Ava's experimental approach tests whether nrf2 binds to viral DNA in vitro using Chip
Test Samples: Viral DNA sequences (three viral promoters) were cloned into plasmids. The color denotes timing of gene expression in the virus lifecycle (red, Immediate early; purple, Early; green, Late).
hypothesis: Nrf2 binds to promoter regions important for early infection steps
DNA agarose gel of Ava’s PCR to confirm the her experimental approach. Plasmids (P) were transfected into HeLa cells using lipofectamine then either treated (T) with MG132 or left untreated (UT). DNA from those samples was used for PCR to detect the regions of interest. She is currently performing the chIP experiment this quarter and hopes to finish by graduation!
Thanks for watching. For more information or questions, please reach out to Dr. April Burch email: aburch@berkshireschool.org