Week 3 (9/13 – 9/17)

Hey everyone. For me, the early part of this week was oriented around the two refreshers. By Wednesday, I managed to finish both of them and upload my results to GoogleDocs for you all to see. During the latter part of the week, I spent time organizing my lab notebook for Friday’s check, preparing and sending off my Midiprep results to be sequenced, and analyzing the results. I just finished comparing and BLAST-ing my results today so I’ll be uploading those to GoogleDocs this weekend as well. As for next week, I’ll be focusing on two major tasks – The first is the target research report, which I hope to beging very early on (if someone knows where the instructions/guidelines sheet is, please let me know!). The second is expressing and purifying my pNIC-Bsa4 + CA7 protein. I hope to finish this by the end of the week and finally be ready to go with some virtual screening and (hopefully) find some good inhibitors. I hope everyone has a great week ahead of them and I look forward to seeing you all in the lab!

MIDI Prep Warning

Hey VDSers,

When doing the MIDI Prep, first make sure that the two tubes you will be using in the steps are two tubes of your assigned protein. I messed up earlier in the lab because I threw away my supernatant, but in hind sight I am happy I messed up. If I would have kept going for the whole 6 hrs I would be doing the MIDI Prep again next week because I was using the pNIC and my assigned vector. Don’t Do That. Both tubes should be your assigned vector. Next, when it says remove your supernatant on the protocol, it mens to remove it into another tube, not in the supernatant waste. Keep the supernatant on Steps 7 and 8. After spinning, which is the beginning of the lab, is where the supernatant would be removed as waste. Lastly, when adding the P1 buffer in step 4, resuspened the cell pellets when you add the P1 in tube 1 and also when you take the solution for tube 1 to tube 2. This is important. When you transfer your tubes, you will not be able to see the cell pellets in tube 2 because its the same color as the solution. Resuspend.

I am letting you guys know so you will not make my mistakes.

Da’Marcus Baymon