Week 6- Third Time’s the Charm!

Anita M -

After making modifications to our protocol, we were able to add the GRASP55 plasmids to the cells with minimal cell death!

We also plated only 400,000 cells this time, again, to avoid overcrowding in the wells. As you can see, compared to the last two trials of this experiment (in my last two posts), our starting amount of cells is a lot less.

This afternoon, I will treat my cells with a chemical called tunicamycin (TM). The TM treatment causes stress in the cells to emulate the stress the cells undergo after a heart attack. Tomorrow, I will begin the process of extracting genetic material to see how our genes of interest (ATF6, AMP, and GRP78) have responded to insertion of the GRASP55 plasmid in the cells.

If everything goes to plan, I’ll have some more images of my cells to share with you guys in my next post!

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    camille_bennett
    Hi Anita, I'm so happy your cells survived!!! Is there a down side to using less cells? What other protocol changes did you make to allow them to survive?
    March 25, 2025 at 10:45 am - Reply
    anita_m
    Hi Ms. Bennett, the only down side of using less cells is potentially getting less genetic material from them later on, but we didn't run into that problem, thankfully. Other than plating less cells, we also skipped over a buffer wash step of our procedure to ensure the cells had a constant supply of nutrients, since we hypothesized that step as another potential reason for cell death and skipping that step won't affect the results of our project.
    March 25, 2025 at 12:14 pm - Reply

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