2096 2097

Date: 2014/2/20

Cells: BL21-AI, 5 ml, induce with 0.15 mM o/n at 18 oC. (cell initially all lysed, only a few survived, then growed to OD600 ~=1 at 37 oC, cool down to ~20 oC, add IPTG to 0.15 mM).

Purification: 50 ul Ni beads

S: cell lysate supernatant, beads elute with 25 ul 0.3 M Imidazole, first elute

S2: beads elute with 25 ul 0.3 M Imidazole, 2nd elute

P: cell lysate pellet, add 8 M urea to dissolve the pellet, add 50 ul Ni beas, beads elute with 25 ul 0.3 M Imidazole, first elute

P2: beads elute with 25 ul 0.3 M Imidazole, 2nd elute

Conclusion: Yeast version of Flp is better than E coli optimized high CAI version