12.19.14 Friday, protein gel and buying presents
12.18.14 Thursday, catching up on things
I didn't get as much done as I wanted today - but I did a bit of everything - read a little of a GFP plasmid paper, poured a 12% SDS-PAGE gel to run on the protein tomorrow, finished lab notebook notes, cleaned my desk, ordered some supplies and sent emails to some researchers. All very important. I really enjoyed being around all my labmates. I love them all so much. Coming home was a nightmare - horrible traffic jam on 78 leading to a 1.5 hr commute. Here's hoping tomorrow I can get TWO gels done - protein and an activity assay! We shall see. :) I also have crazy amounts of shopping and other things to finish before next week.
12.17.14 Wednesday, protein dialysis, mystery of the pink buffer
To complete the protein dialysis, I wanted to do a few more buffer exchanges, once an hour. Remember those office clips? When I walked into the cold room, I found that BOTH protein buffers had turned pink! I was alarmed. What did this mean? It took me a few minutes to realize that our office clips had actually rusted out, turning the buffer pink. >:| Of ALL the things! That meant, of course, that now there were tons of iron ions floating around in our protein too - and who KNOWS what that would do to it. I've read that iron doesn't inhibit our enzymes activity though. I changed out the pink buffer and tried to do more dilutions, but I'm not really sure that I ever got rid of the iron. I doubt it. |
I came in at 8:30 am instead of 7:30 am, but I felt totally exhausted from the previous day's 14.5 hr + commute work day. Very unusual for me. In the afternoon, I realized I forgot my lunch from home, and tried to eat my leftover Chipotle from the day before instead, but my body said no. I couldn't eat it. I ate half a yogurt. Then, I had a headache, was exhausted and felt really nauseated. I showed Sam how to freeze some protein samples, and then expected I'd need to freeze 160 additional tubes myself. That's a lot of tubes to label. I didn't think I could do it - I was feeling that sick. I asked Kevin and Kornelia to help me label tubes, if they wouldn't mind, and they said sure! But first, i realized I should check my protein concentration to make sure I COULD freeze all of it straight up. I was hoping for .8-1 mg/ml, but it was .2 mg/ml. I decided to risk concentrating it. Before, when I tried that, my protein would precipitate, but that was because it was 1 mg/ml already and I didn't know it (though technically, it *should* be able to concentrate more than that - must be the buffer I'm using). When I tried concentrating it, the protein precipitated - both of them. :| But, I checked the concentration of what was left, and it was .9 mg/ml! ... Weird, huh? Well, at that point, there was only enough protein for about 70 tubes instead of 160, so I was like, great! I quickly drew those out, labeled them briefly, aliquoted the protein, flash froze it in liquid nitrogen and did preliminary cleanup.
I expect that protein to be no good, but hey, I want to know what it's weight is and test it for activity anyway, you know? Sam said, maybe iron is what we needed all along and we didn't know! Ha ha - you never know. :) I went out to eat at 5:30 pm with classmates for an end of the semester dinner - it was so nice to see them all and talk to them. I especially had missed Wallace and got to hear how he was doing. Additionally, I got to meet Morgan's husband Tyler! That was awesome.
More Rusted Buffer and Us Second Years Hanging Out at Slice and Pint at 5:30 - 7:30 pm
I expect that protein to be no good, but hey, I want to know what it's weight is and test it for activity anyway, you know? Sam said, maybe iron is what we needed all along and we didn't know! Ha ha - you never know. :) I went out to eat at 5:30 pm with classmates for an end of the semester dinner - it was so nice to see them all and talk to them. I especially had missed Wallace and got to hear how he was doing. Additionally, I got to meet Morgan's husband Tyler! That was awesome.
More Rusted Buffer and Us Second Years Hanging Out at Slice and Pint at 5:30 - 7:30 pm
12.16.14 Tuesday, protein purification and box extravaganza!
So, there's a reason you didn't hear from me on this day - I got in at 7:15 am and left at 9:54 pm! I was doing prep work until 10 am, at which time my student Sam and I were supposed to start isolating and purifying our protein. However, he had just finished finals and overslept by accident. Meanwhile, we also had to clean a mess of boxes downstairs at the loading dock. Sam came and helped with the boxes with me and Edward, and by the time that was finished, it was noon. THEN, we started the purification. At that point, I knew it would be a late day. We had fun though! At the end, we were putting our ~7 ml volume of each protein into dialysis tubing and letting it sit inside of another buffer to let the imidazole (small molecule that we had to use to elute our protein) to diffuse. Imidazole is bad for proteins - denatures them and stuff. However, we couldn't find the plastic clips. So, we made do with office clips and felt victorious. ;) Our protein would dialyze overnight. I told Sam he'd experienced a realistic, unpredictable day of experimenting - especially with protein - which can make for random late days. He said he didn't mind and was enjoying it. I got home around 10:35 pm, talked to mom and dad a few minutes, ate a dinner of tacos and went immediately to bed at 11:15 pm.
12.15.14 Monday, attempting to do MALDI and failing
Today, Josh popped in in the morning, carrying plexiglas and asked if I wanted to go down to the machine shop and cut it with him. (...) I said "sure!" To see that, check the science blog.
Today, I prepped a last buffer for protein purification tomorrow. Sam and I expressed P.horikoshii RtcB and archease last Thursday and stored it in the -80C freezer. He had finals today, so I wanted him to get to see the purification - thus, we're doing it tomorrow. I also tried to do MALDI, but that failed. I could never find the appropriate protein MALDI plate. Hopefully I'll find it soon and try that again on Wednesday when we freeze the protein.
John got the leads to my car battery replaced (after 45 min of wrestling with the bolt on the positive terminal) and it works fine again! Thank goodness it was only the battery and thank goodness for John's handyman-ship.
Today, I prepped a last buffer for protein purification tomorrow. Sam and I expressed P.horikoshii RtcB and archease last Thursday and stored it in the -80C freezer. He had finals today, so I wanted him to get to see the purification - thus, we're doing it tomorrow. I also tried to do MALDI, but that failed. I could never find the appropriate protein MALDI plate. Hopefully I'll find it soon and try that again on Wednesday when we freeze the protein.
John got the leads to my car battery replaced (after 45 min of wrestling with the bolt on the positive terminal) and it works fine again! Thank goodness it was only the battery and thank goodness for John's handyman-ship.
12.14.14 Sunday, gearing up for the week and Christmas concert
Today I went to church, but John stayed home because he didn't sleep all night. We ate tacos at the Petree's house. My battery died in my car, so John is getting a new one later today. Dad, mom, James and I are going to this concert at Dr. Stanely's church. It sounds really nice. Mom and dad are also going to see White Christmas in the theatre later today. After the concert, I'll be blogging and looking up science ideas.
12.13.14 Saturday, a day of rest for us
Today, John and I played Dragon Age 3: Inquisition, helped each other with it and later played multiplayer with Brian and Matt (our Warmachine friends). Mom and dad got a BEAUTIFUL 11 ft Christmas tree in the evening and put lights on it! (See pictures here). Also, look at the date! It's sequential! Last time that will ever happen.
12.12.14 Friday, James's birthday and lab cleaning
Today was James's birthday! He turned 25. I slept in, having been exhausted from staying late in lab on Wed, and got to lab late, missing a nanoparticle seminar. I felt really badly about this. Our lab had lab cleaning - I was really pleased with what I got done. I cleaned out all my tubes in the cold room, put in the rest of the items into the inventory and cleaned the biological area - even getting the melted rubber rings stuck to the counter that the little microfuge tube heaters leave. I left at 6 pm - later than I had wanted - and got home at 7:20 pm. James opened birthday presents and we had pot pie. (See pictures here). I got him Shadows of Mordor for the PS4 but he decided to trade it out for the new Call of Duty. I have to say - it is far more realistic than the old ones - which I am not sure how that is possible.
12.11.14, Thursday, protein expression w/ Sam and lunch w/ Keon
Today I expressed P.horikoshii RtcB and archease with Sam, my student. We got a late start, so had to stay until almost 9 p.m. I ate lunch with Keon, and it was really great. We both got to encourage each other in different ways and talk about the Lord. Keon prayed a beautiful prayer. I'm looking forward to the new year and getting even more organized.
12.3.14 Wednesday, the lost day
We'll see.
12.2.14 Tuesday, chemical transformation of our plasmids
Ran around crazily, seeing if I could do a transformation, and finally achieved it! We got the archease plasmid from the Raines lab - so exciting. Worked on making SOC media, LB and other things. More updates soon. Things are finally getting underway!
12.1.14 Monday, preparing protein expression buffers
Worked on getting buffers made for protein expression with my student Sam.