July 13th - 24th, 2015, My RNA has died a lot
July 12, 2015, Sunday, tired, making to-do lists and hanging out with John
July 11, 2015, Saturday, running lots of errands, lots of errands
John plays in Warmachine tournament. Meanwhile, John went to an all-day tournament for Warmachine in Marietta. I drove up there after my errands and a nap. He bought me a brownie Sunday from Mazzy's here they ate, which was delicious. But come 8:15 pm I had to go back home because I was so tired. We ended up going to be quite late. I just read until he got home. John won his first two games, but played Brian White in the third round and lost. This is acceptable though because Brian is a world-champion Warmachine player. However, even so, he was one of eight people who made it into the second round on Sunday.
July 10, 2015, Friday, running two gels
Experiments are slow but accomplished! My experiments took way longer than I expected to setup, because while aliquotting out the reactions, I ran out of both of my small stocks of RNA strands, so had to make fresh and Nanodrop them - never a fun prospect. Finally, I got done around 8 p.m. Also, I left my phone at home by accident, which worried John. But I sent him email updates, which worked.
July 9, 2015, Thursday, freezing protein
Grace did a pH experiment. Meanwhile, Grace ran a gel of two different pH's to test how our enzyme worked in them both. She did a great job - it showed some pretty cool info! I was impressed. I wasn't sure what would happen with that one.
July 8, 2015, Wednesday, purifying protein
July 7, 2015, Tuesday, really busy! lab meeting presentation + gels
July 6, 2015, Monday, feeling frustrated
I came home and jogged -- now it's pouring rain. Looking forward, in some ways, to presenting tomorrow. Then, finishing up protein expression, and untangling this unruly gel.
July 5, 2015, Sunday, Preparing for next week, excited about experiments to come
Thus is today! More next week! I'm very excited about the experiments to come!
July 4, 2015, Saturday, Happy July 4th!!
Glow Globe. I was really excited about that glow globe we launched. We saw them last year, and we really wanted to try them out. So, this year, we launched our own!
Big fireworks are scary. Also, the big fireworks we launched were scary as crap. HOLY COW! They were really loud. After my brother launched about seven of them, I'd had enough. They were moderately terrifying. It'll take a while to get used to those. Our neighbors were also launching their own gigantic ones, some of which were bigger than ours. We got quite the show!
July 3, 2015, Friday, John comes to work with me!!
We did an experiment when we got back, but sadly, it kind of bombed. I'm not exactly sure why, but I'm going to give it another shot Monday. When we came home, for some reason, I was really tired and took a nap until 7 p.m., when dinner was ready.
Having John there all day was such a treat! I loved having him at work. I showed him the -80C freezer and everything, and a few things I was doing with experiments.
July 2, 2015, Thursday, Protein purification insanity post-poned!
What I did instead: Expression and RNA labeling. So I labeled some RNA for an assay later. And I expressed my enzyme (half of what I had planned) but did not purify it. I froze it for later. I just need to do the purification and dialysis in one day. I can split it up at the expression.
Peaceful awesomeness. It was a very peaceful, fun and awesome day -- one of those days that's totally normal, and one is doing one's work, feeling happy and vaguely excited and satisfied with life. I got lots of packages from the stockroom, re-read a Shuman paper and made notes, wrote up Dr. Weinert's and the Lutz lab people's thoughts, wrote my own notes, enjoy a lunch of perlo and bread ... ahh. This is the life. I love no class and just doing what I love best: research and thinking. Thinking and research.
Tomorrow will be better! Tomorrow will be even more awesome because John is off work, so he's going to hang out with me at Emory! I'm so happy about that!!!! Ha ha. What a great week. Until next time!
July 1, 2015, Wednesday, Preparing for tomorrow's day of insanity!
Talking to the Lutz lab about protein expression. I talked to Pravine and then Samantha (Sam) a long time about protein expression. My yield was a little low when I expressed it with Grace, so I hashed out with them some ideas of how to increase it and how to concentrate it. I had a lot of fun talking to them and they had some great ideas.
Synthesizing more gold nanoparticles! Today, I synthesized 100 ml gold nanoparticles for the third time. This time, I tried to do all the tricks I'd learned the first two times. Kevin wasn't even here, so I did this one totally by myself! Yun and her undergrad Nick watched. I really hope it worked this time - no fused particles AND most of them the same size - that's what I want.
Preparing for protein expression. Tomorrow, I'm going to express protein and purify it in the same day - the experiment of DOOM of DOOM!! This is one I really am glad I only have to do it once a year. It takes from about 7 am - midnight. I have to do it like this to try out the tips Pravine (Lutz lab) mentioned to me. I'm going to try to concentrate my protein in a unique way -- I think it will work, based upon the tests I ran on it this past week. I'm REALLY EXCITED about it! Maybe I can finally get concentrated protein this way!
Technical jargon - what am I doing to concentrate it:
My protein precipitates in Amicon filters when I do buffer exchange on the 7 ml of ~.2-1 mg / ml protein I get. So, I typically dialyze it overnight and freeze the resulting batch as is. Pravine said I could leave the dialysis bag in open air to let the extra water evaporate slowly (keep it in cold room though). That's a genius idea. I think it will work, because I tried to concentrate my protein in the speed vac, vacuuming off the excess liquid, and it worked! The protein concentration got up to 2.1-ish mg / ml! Normally, it would precipitate out. I'm wondering if it worked because the salt and buffer concentrations increased as well, since I'm just removing water, helping the enzyme stay happy. Therefore, this method should work too. But, I wanted to dialyze the protein, then leave it overnight to dry -- I don't want to risk it overnight and not flash frozen any longer than I have to -- thus everything has to be done in one day. To achieve that, I have to do the one day expression / purification insanity. But I think it'll be worth it!!
Visiting James. While going home, I saw James driving up to visit his girlfriend Elizabeth (she lives right next to Atwood chemistry). So I visited with them 30 min and met her dog Milo. That was really hilarious!
And tomorrow is a NEW DAY full of more AWESOME! Until next time! Same bat time! Same bat channel!
Pictures of gold nanoparticle synthesis!!