End of the first week.

I mainly worked on preping RNA from animal tissues this week, although I did learn how to spec RNA as well(detect how much RNA was extracted). The RNA comes from the tissues that I discussed in my last post. I extracted RNA from the Fat Body, Ovaries, and the Carcuss. RNA is extremely unstable, so making careless mistakes can contaminate your samples.

Preparing RNA is fairly simple. First put on gloves(verrrry important). Next you take a buffer(RLT) and pipet 10 microliters Beta ME(macryptoethanol?) for every mililiter of RLT buffer one uses. Then 350 microliters of this buffer is added to the sample tubes. Because the tissue is, well, tissue, you use very small pestles to carefully crush the tissue to make it easier for the Buffer RLT + Beta ME to break open the cells. After the tissues have been crushed, you put them in a centrifuge for five minutes. This takes all the tissue and presses it up against the wall of the sample tube. Next, you pipet the remaining solution into a labeled "Sample Tube". After that you prepare QIA cube so it can do the rest of the work for you.

QIAcube does many of the RNA prep steps for you. It centrifuges, pipets, and filters to complete the remaining RNA processes. However it is very important to understand what it is QIAcube is doing in order to properly troubleshoot it in case something goes wrong. One of the most important steps of preparing QIA cube is making sure that the chemicals(two buffers, RLT and RPE with ethanol; RNase free water; and a 70% ethanol solution) are at proper levels. On tuesday, after Anam had left, I asked Jenya(Another extremely intelligent individual working in the lab) if I could simply add more of the RLT buffer(as it was low) to the RLT that was already in QIAcube. She said yes, and I did so. I began QIAcube again, and again I got an error saying that RPE + Ethanol was low. I retrived the RPE buffer from the RNeasy buffer box and poured it into the RPE buffer, without adding ethanol.

I know now that ethanol needs to be added to the RPE(to be specific, 44ml of ethanol needs to be added for every 11ml of RPE). Because of this mistake 10 samples are possibly unclean, and therefore unusable. I was reassured that mistakes happen, and mistakes are how we learn(by Anam). It was then that I realized that working in this lab would be a development of not just my lab skills, but my work ethic and my maturity.

On friday I attended a seminar on a research paper that another lab had done that was relevant to our own. Every week a member (or in this case members) gives a presentation on said article. I was surprised that I was able to understand some parts of the presentation, as it used very specific biological terminology, some of which I had learned from Anam earlier.

After the seminar Kendra took me up to the 8th floor and taught me how to do RNA specing. You wipe off the surface of a metal arm and a extremely small platform that has a hole in it. The hole allows the liquid to pass down into another chamber, where a lazer determines the nanograms per ml of RNA in the liquid. I examined three samples after she demonstrated with three samples herself.

I am looking forward to learning more lab techniques and more about mosquitos. Hopefully I can prevent future mistakes with increased caution and by asking when I am unsure of how to do something. All in all, this week was an extremely sucessful learning experience, as I am sure the weeks to come will be.