This is a day for celebration!!
Chronicling the Second Year: Before the Talk
Nervous stress. After I had setup my laptop and fixed up the laser pointer, I had twenty minutes left to wait. So, I went back to Khalid's office and asked if I could keep talking to him until the time, to keep from being terrified. And he let me! So nice of him. He promptly talked about nothing in particular, which is very comforting and he is good at. He got rid of those toffees in his office and has snickers now! I approve.
Photos: Gokul with the tribble
The presentation ... dun dun dun ... !
The tribble. I remembered my tribble at the last minute and ran to get it. It's a fluffy round alien from Star Trek that is supposed to multiply rapidly, like rabbits. It makes soft purr / trills when it's happy and an angry screech at Klingons, which mine does both. I gave it to Gokul to hold for me. He found that amusing. It was important for the tribble to be there to "witness" things. I started bringing it during the summer I rotated, and would bring it to lab meetings regularly. It acts as my mascot. Apparently, Dr. Lutz was giving Gokul and the tribble a lot of strange looks and Gokul said no – he wasn’t distracting me – I had *wanted* him to hold it.
Starting off. Wow that was scary! Starting off, it was about as I expected and didn't seem so bad. But then Dr. Lutz asked the first question on my second slide. And I couldn't answer it perfectly. AND THEN, things got terrifying. I felt like I was naked in a room of experts saying things in which they could obviously see flaws. I talked not as loudly as I had practiced. And I absolutely did not look at my committee – or Khalid – as I didn't want to see what he might be thinking. I was terrified of them. I just watched Gokul and the tribble, which was a nice center point to keep me sane. And the rest of my labmates looked vaguely bored, which was somehow comforting.
Difficulties. At one point, I lost my train of thought and really bumbled, completely forgetting my last point about why RNA editing was more advantageous than DNA editing. Worst point in the ENTIRE talk. I thought I might fall apart, but I just gave up and skipped it and slowly started being more calm again. The committee members asked more and more questions. I was horrified, that while discussing my strategy to make my nanozyme to splice RNA, Dr. Lynn pointed out a completely amazing flaw I had never noticed before! I was like ... WHAT the heck! AHHHH!!! And I said he was right – that was a flaw.
Feeling terribly not confident. Some of the questions I answered but didn't feel confident about. My answers were more vague than I would have liked. I should have read over my papers on the background more carefully a few days before, just to make sure I knew the details better. *headdesk* "More literature reading! Should have done more literature reading!" I kept thinking. After I had bumbled on that slide and Dr. Lynn pointed out that flaw, I felt doomed, and decided – well, I would do the best I could and whatever happened happened. So, I started caring less about how stressful everything was and just kept going, trying to sound confident, in spite of being in front of an audience of experts. Not easy to pull off. I'm so glad I had a lot of people praying for me.
Brief audience afterward. After the presentation was over, I stayed behind and they asked a few more things, but said they’d asked most of their questions during the presentation. Then they told me to go, for their closed meeting.
Emerging to talk to friends: I left, feeling surreal that I was coming out, as I’d seen other second years do – now it was me. I wanted to cry. I thought I’d done terrible! But Kevin, Gokul, Yuan and Rolando were there and said YAY! And I gave them all hugs. They said, no I’d done well with the questions, and Yuan said that Roxanne sitting next to her kept saying that I was doing awesome. They said that the committee wasn’t being vicious – they were just genuinely curious. This was by no means a mean second year – they’d seen other people get ripped apart for real.
The closed door session: after presentation comments
Initial comments. They had me sit down between them all, which felt somewhat intimidating, but they didn't seem unhappy. Dr. Lutz did most of the speaking and said that I had done well fielding questions, which really surprised me. He commented on this, saying that I was giving him a "whatever" look. Dr. Lynn said it was good the questions didn't cause me to lose my train of thought. And Dr. Weinert said, knowing how terrified I was of giving presentations, I had improved a lot in the last year. But all of them said that I needed to look at them more while I was speaking – only too true. Dr. Lutz also said there were a number of very challenging problems to my work and I’d picked a very difficult project! But, that doesn't necessarily make it bad – there could be interesting things that come out of it, even if it’s not exactly what you set on.
Feedback for the project. They had lots of great feedback! And they didn't pull any punches. They said RtcB was the weak link in my chain and I needed to ditch it. Dr. Weinert said that RtcB was not, in fact, the only enzyme that could do this ligation. There were others, and I should screen them to look for ones with higher activity that don't need cofactors. There were some other very interesting ideas thrown around. It really helped shape my thoughts completely differently about my work. I hadn't realized there were other enzymes.
The plan going forward
- Acquire E.coli archease to see how it's activity affects E.coli RtcB. I'm not sure this is completely necessary.
- Screen other enzymes for ligation activity that's higher.
- Build DNAzyme nanoparticles that can cleave in two places in a non-functional GFP transcript and see if a cellular ligase, rather than an attached ligase, can do the ligation step.
I'm excited! There is now a LOT to do. Unfortunately, most of it will take a lot more planning before I'm ready to do major experiments. So, primarily right now, I'm focusing on editing my NSF proposal for the deadline on the 30th and getting ready for a presentation in Dr. Lutz biotech class on the 17th.
Thus is the news so far! God is good! I yet live!