Student Research at Duke

soooo i was making minipreps at 7 pm friday night, and i was supposed to wash my DNA with one buffer, centrifuge, and then add another buffer to dissolve the DNA in a solution. Sadly, I washed half of the minipreps with the dissolving buffer, and so half of my DNA was promptly sucked away through the vacuum manifold.

well anyway, as Tammy and I were making a gel for my remaining DNA, we discovered that we both like 80’s music! and so does Eric..so maybe we’ll have an 80’s party some time while we sort mutated fruit flies.

the fly roomrnrnthe other side of the fly room, with a couple of my lovely coworkers!rnrn this refrigerator is FULL of flies! it’s about 3x as big as what you can see in the picture. I actually have a PHOBIA of bugs. but you gotta do what you gotta do, right?rnrnThat’s me, annealing some DNA. Luckily I will be doing more biochemistry in my project than fly work!rnrn I hope nobody noticed me posing with the micropipette in front of my computer using my webcam.rnrnWell anyway, I’ve been working on a UV cross-link so far. I started with an RNA sequence, then i designed mutated versions of the sequence and ordered single stranded DNA oligos online. They came in the mail as tiny specks of powder in little vials. I resuspended the oligos in a buffer solution, then annealed the matching strands by boiling and cooling. At this point I grabbed a plasmid containing the ampicillin resistance gene and a promotor sequence and digested it with a restriction enzyme, then sent it through gel electrophoresis for purification. Finally I ligated my new double stranded DNA strands to the digested plasmids and then transformed the vectors into bacterial cells by heat shocking everything. Last but not least, I spread the mixtures on agar plates, and hopefully later today i will be able to pick off some bacterial cultures that have cloned my DNA and produced mRNA for me! All of this involved lots of temperature changes and incubating, and so it has taken all week since i got the oligos in on monday.

i’m ordering oligonucleotides tomorrow…GATTACAAAA