Experiments This Week
*Week after talking to Dr. Weinert (previous Friday)
Monday – prepared buffers for protein purification, washed Ni:NTA columns, made stocks of the hybrid DNAs in 10 mM Tris-HCl (phos 1, sub 1b, 20mer) and froze them at -80C. Made overnight of Keio delta rtcA::km/pQE70-rtcB::amp
Tuesday – Finished making more of the buffers for expression (buffer B1). Finished buffer C’s w/o the DTT added. Attempted o/n expression but the Weinert incubator was full.
Wednesday – rtcB protein purification of Raines protein from previous Friday, according to Dr. Weinert’s instructions. Concentrated in Amicon filter. Saw precipitation. Yvonne showed me how to flash freeze my aliquots in liquid nitrogen.
Thursday – July 4th – did not come into lab
Friday – Nanodrop of rtcB protein; concentration indicated to be ~0.016 mg/ml. Poured 18% RNA gels using silicone grease on the edges to seal. Discovered had to allow the grease to set or the plates would skew apart at the bottom and leak. Important not to touch the setup after it’s in place.
Purchases for the week = $212.33