Monday, April 27, 2015, 5'-end labeling my RNA with new reagents
Since I last wrote, I was able to incorporate aminoallyl-UTP into my RNA. I attempted to label it with Cy3b and was partially successful. However, I also saw some degradation and the reaction was not optimized. I think that I've determined the degradation is probably from contaminated DMSO - I used the lab's stock - which is used a lot by everyone for DNA work. I bought my own, and today, tried part 1 of the experiment to label my RNA 5' ends with 5-iodoacetamidofluorescein. Wish me luck!
I will get to an actual update sometime soon. Partly, I have been scarce lately due to a lot going on and to adding gaming with John back in as a hobby, rather than only blogging. I was feeling restless for something relaxing to do, and this has really helped. Unfortunately, however, it has also made writing more scarce. Class has ended, so that helps a lot. I hope to write more and not just technically. Things will happen slowly - I'm still working out what I want my summer schedule to be. I may schedule a day for writing. However, it is late, so I can't do much today.
Thursday, April 9, 2015, My modified UTP came ALREADY!
Photo Notes: Also, in the morning, I found this giant slug on the pavement. It was so cool. I love giant slugs. I put him in the bushes of course. There were pretty flowers on the way to class, so I took a picture of those too. Maybe you like them better than the slug? I don't know. It's a toss up to me.
Sam had a test today, but still came into lab after it, and helped me pour a gel. I was so grateful. Thanks to him, I could actually get things done at a reasonable hour. AND THERE WAS NO REACTION IN THE NEGATIVE CONTROL TODAY!! That's a super good thing, just sayin'. I have no idea why there's been autoligation / reaction without enzyme earlier, but there has been and it's annoying as heck.
I also spend a portion of the day pouring over notes on NHS ester dye coupling reactions and talking to Yun about them. I think I'm going to try Cy3b first, because that's what we have available in the dye box.
In class, I ate my lunch and a quarter of a huge chocolate rabbit. The professor talked about adenoviruses. I was exhausted all day - I think it was the heat. But I finally got Claritin so my eyes aren't *as* itchy. Pollen count of 6,125 today! Ok bye, got to sleep!
Wednesday, April 8, 2015, Figuring out RNA labeling
And I'm so encouraged! I think it'll work! I found exactly what I needed: aminoallyl-UTP, that goes right with the transcription kit I'm currently using. I'm so pleased! I ordered it and new supplies! I can't WAIT til it comes in and I can try it out!
Meanwhile, I finished labeling the 5'end of some transcribed RNA that Sam made with a FAM. I'll have to see if it worked and the RNA didn't degrade tomorrow. It seems as if the RNA pellet was the correct dyed color at least, which is a good sign. I also autoclaved pipet tips, checked the calibration of Kevin's pipets that I always use (they were great) and am currently baking some glassware and spatulas in the oven overnight so they will be RNase free.
For lunch, I ate with Brandon Greene, Amanda (Heaven lab) and Zed (Lian lab), taking Dr. Wang out to lunch. Kira asked if we'd host him and we agreed. He was giving a private seminar to faculty only. I felt kind of privileged to be asked to host him with the rest, though honestly, someone in the Hill lab should have subbed in. He works on batteries and nanomaterials related to those things. I tried 4 cheese ravioli at Amanda's suggestion at Saba for the first time. It was, thankfully, quite good! I needed something good. Before lunch, I drank peppermint tea and immediately got an awful headache for some reason. I decided maybe my body was revolting against a non-caffeinated drink.
LOTS TO DO!
Other news, in real life
Tuesday, April 7, 2015, My parent's 30th anniversary and experiment bust
Class was LONG. That's one thing I remember. And the professor talked so fast it hurt to listen. I wanted to absorb what he was saying, but I couldn't at that speed. He was talking about Epstein-Barr virus (mono) which I've had, so I was really interested. It's actually kind of a fascinating virus and indirectly related to the enzyme I'm studying right NOW - which amused me greatly.
The experiment was kind of a bust. I forgot to do something to it - but if I had - it would have taken too long almost to be worth doing - so I don't know what I should think about that. But oh well - it was useful-ish.