Happy to announce that my three+ months in Panama has resulted in one tiny tube containing 60 microliters of data!!!! That’s basically one fat drop of liquid containing all of the DNA I sampled in the ocean.

This is literally all I have to show for my work right now.

To visualize the craziness of how far we’ve come with molecular tools and sequencing, I’m including what all it took to get this tiny tube:

1) Collect 144 samples from the ocean every day for three weeks and filter them on the boat for DNA on a small circle of material that gets put into a falcon tube.

Look at all these places I visited! Not included are the boat docks I also sampled at around the main island.

4) Dilute your extracts, set up hand-sized (!) PCR plates, and target the part of the DNA sequence you want (I wanted a region of the DNA that characterizes fish/metazoans from each other and not everything else) and add on primers that are unique DNA ID’s for later analysis. Clean up those plates with all of your samples….very gingerly.

4) Combine samples all into one row of another hand-sized PCR plate. Remember to breathe since everything still looks like there’s nothing in it but you know there’s something in it.

5) Ligate (add) on adapter (more DNA) sequences to further “multiplex” or differentiate your sequences from one another and….combine everything…everything you’ve worked so hard to not cross-contaminate, into one tube. Adding this photo again to emphasize how freaking tiny this tube and volume of liquid is.

Rejoice if the gel electrophoresis and DNA quantification tell you what you did was correct (this was also done throughout all of the steps mentioned above).

THIS EGGHEAD IS SO HAPPY!!

Working in a molecular lab for the first time ever on my samples was so nerve-wracking! I have never lost trust in myself so much as I have here. Staring into the tiniest amounts of clear liquid, going insane over how meticulous I needed to be in tracking my samples from one tiny well to another, and feeling satisfied with what I have accomplished at the end of each day have all taught me important lessons about my self-perception and confidence. It’s a mental game, one that I went into head-first. Every single thing I did (other than running a gel) was my first go at a protocol I had never done before on my samples…no trial runs. Plus, the days were melting away and I could feel the pressure of my departure date looming upon me. I cannot tell you how many times I stared into small tubes of clear liquid with my heart in my stomach and anxiousness in full swing, when I doubted myself and my ability to pipette correctly and accurately. I’m honestly quite shocked that my samples worked (well!) and that I have DNA worth sequencing.

If you don’t trust your samples and don’t trust yourself, you’ll never get a project done. Believe in your work and your ability to execute it. I’m still not confident in myself or my work (yet), despite the numbers and gels telling me so. But just because I can’t see it, doesn’t mean it’s not there. I guess that can apply to a lot of things.

Anyways I just wanted to express my RELIEF and GRATITUDE that I am done with what I set out to do in Panama. I board my flight home in three days!