Tuesday, May 31st: Dad's birthday; I grew E.coli cells
I grew my cells today overnight, hoping to have a nice crop of cells in the morning. I also celebrated my dad's birthday briefly, and got him a few silly gifts: an Atlanta cup from Starbucks, a Sherlock Holmes puzzle game and a cosplay book - it had nice pictures. James and Elizabeth got dad this sparkling lemonade - I tried some - it was actually quite good.
Monday, May 30th: Memorial Day; Autoclave broke
I tried to go in to work on Monday - prepped some solutions for the week etc, but I couldn't autoclave my media for more chemical transformations and culture growth. SO, it was a bit of a bust of a day. I left early, went home, and did lots of laundry. Then in the evening, we all went out as a family for Dad's birthday (which is tomorrow) - Dad, mom, me, John, James and Elizabeth - at the Bonefish Grill in the Avenue. I had Norwegian Salmon, and it was actually really good.
Friday, May 27th: Meeting with Khalid
Today is my weekly meeting with Khalid at 3:30 pm. I'll give him an update on what I did and what I intend to do next week. Here is that update:
Isolating the tRNA that included my splicing substrate failed, though I was able to isolate the plain tRNA by itself. I think I may be able to fix the problem, in isolating the tRNA with the splicing loop: translation - I want to repeat the experiment a few more times to see if I can get it to work.
Isolating the tRNA that included my splicing substrate failed, though I was able to isolate the plain tRNA by itself. I think I may be able to fix the problem, in isolating the tRNA with the splicing loop: translation - I want to repeat the experiment a few more times to see if I can get it to work.
Thursday, May 26th: RNA degradation by alkaline hydrolysis
Unfortunately, today, I made a classic mistake, while trying to isolate total RNA from some E.coli. I put them in too much base, allowing them to degrade into gigantic useless smears. At least I know what happened and how to fix it. However, it was entirely frustrating. Lost some good info in those cultures. I'm going to try repeating it next week.
Tuesday, May 24th: Cells didn't grow today
Today, I was growing E.coli cells expressing a plasmid with a tRNA construct that contains my splice loop - the loop I am attempting to splice. They needed to grow a lot so I could isolate the constitutively expressed tRNA. But alas. They did not grow.
I put in some more cells. These have a different antibiotic marker - Amp rather than Cm - which I've always had better luck with. Hoping they grow by tomorrow. I also did some house cleaning maintenance type experiments and read two articles. See below.
I put in some more cells. These have a different antibiotic marker - Amp rather than Cm - which I've always had better luck with. Hoping they grow by tomorrow. I also did some house cleaning maintenance type experiments and read two articles. See below.
Biomimetic RNA-Silencing Nanocomplexes: Overcoming Multidrug Resistance in Cancer Cells
I really like this article. It discusses a nanomachine that can mimic the RISC complex, which is a fancy way of saying, it chews up RNA very selectively inside cells. Researchers used it to knockdown levels of a transporter that lead to drug resistance. Knocking down this transporter meant the cells were once again susceptible to the drug!
I also re-read a Mirkin paper describing how oligonucleotides on a nanoparticle could determine how cells took it in.
I also re-read a Mirkin paper describing how oligonucleotides on a nanoparticle could determine how cells took it in.