Full day

Today has been quite the full day.  I'm exhausted.  My morning started at 6:30 with transferring products from a lot of little tubes into plates, since they are easier to transport.  I then commenced the drive to Greenwood, where I ran three PCR reactions.

At the GGC, you sign up for PCR slots so that you can get your reaction ready and immediately put it in the machine.  Otherwise the reaction may be ruined as the ingredients could prematurely take their course or denature or degrade.  And you should always check to make sure that the slot you signed up for is open.  I did just this, but when I brought my first two plates up to the machines, someone had decided that they were going to take my slot.  Luckily some others opened up such that I could utilize them.  After putting those in the machine, I went to prepare my third plate.  I came back, and someone was in my spot AGAIN.  I had to go to the back and use the older machine, which I hope is still working properly. It's so important for everyone in a lab to be organized and coordinated; the lab will not run smoothly otherwise.  Working at the GGC this summer really opened my eyes to that.  Having been exposed to only classroom labs before that, I had no idea the organization that had to be in place.  In a classroom, the teacher basically directs all the procedures.

I also got four plates ready for submission to the sequencer.  The process takes a little over an hour to complete, so I could only get two done before 3:00, when the plates are taken over to the building with the sequencer.  I had the option to leave around 3:30 and just get my next two plates ready for sequencing next time, but since I am on a huge time crunch I stayed until 5 to get those plates ready.  It was one of the first times I did the processing sequence by myself.  I never knew what to do without instruction before, but after having performed all the steps by myself once I knew what to do.

I looked over the first two exons I put in the sequencer. Rather, I looked over 140 sequences of DNA and compared them to the original exon sequences.  This process took awhile.  Any anomalies I saw, I highlighted and printed out.  I then left these for Melanie.  One, or both of us, is going to look to see if the base changes could change the amino acid for the protein, and then if that change is likely to be detrimental to the proteins' function.

I dropped some stuff off in the lab upon returning to Wofford; new polymerase, buffer, dNTPs, etc. have to be frozen/refrigerated.  Tomorrow I will continue working on amplifying and exosaping the samples for each exon.