We‘re Not Yolking Around! – Week 4 Update

Okie dokie. Well, this week we decided to go out on a limb to see what direction to take our project. You may (probably not) remember that I ended my post last week with the idea that we are looking to incorporate another age of embryo into our project, either much younger or much older, in order to have a better chance of getting interesting and more exciting results from our RNA extraction. Dissecting older embryos is probably the more reliable/easier option since we would not have to change our protocol all too much…but what fun is choosing the easier route? Thus, this week we embarked on attempting to extract RNA from younger, almost non-existent, embryos.

Extracting RNA is nothing to yolk about!

Extracting RNA is nothing to yolk about!

The youngest stage we had been dissecting previously, was 4 days old. We had chosen this stage because the embryo at 4 days old is visible and tricky, but not impossible, to extract. Going any younger than this was actually impossible, or so we thought. Instead of extracting the embryo from the egg and then running the RNA extraction with only the embryo, we decided to try using a whole egg and extracting RNA from it. There is research to suggest that there is RNA in the yolk and albumin of the egg, so we thought that even if we didn’t get the RNA directly from the embryo, there should still be RNA in the other components inside the egg and getting results from that would be equally interesting. The problem is that this isn’t a very well studied area. There is no protocol to follow and no kits to use, so we basically spent the week trying different techniques, all with the goal of extracting RNA from an entire egg.

We practiced some separation techniques with regular chicken eggs as to not waste any of our precious zebra finch eggs, we tried running the egg through our regular embryo protocol, and we even tried a nucleic acid precipitation (think back to middle school when you extracted DNA from a strawberry), but to no avail. The NA precipitation was pretty cool because we could see the pellet of nucleic acid accumulate at the bottom of our tubes, however it was such a small amount, and most likely contaminated as well, so unfortunately it did not show up on our gel 🙁

Research is full of trial and error (emphasis on ERROR)

Research is full of trial and error (emphasis on ERROR)

It’s been a week of experimentation and many trials (and tribulations), a key step in any research project, but we have reached Friday and decided to turn our focus to the older stages. We are not giving up on the whole egg extraction quite yet, however we are putting it on the backburner for a little while, so that we can get enough reliable (older) samples to collect data from. But there is potential in researching yolk RNA and it’s definitely something I would love to come back to in a few months – but for now, wish us luck as we take our project in a step in a new (and hopefully more straightforward) direction!

Comments

  1. Jacob Warner says:

    Wow! What methods seemed to work the best? How did you develop your methodology?

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