The next day, cells were diluted and transfected at a density of approximately 3–4 × 106 cells per milliliter. Transfection mixtures were made by adding the following components: maxi-prepped DNA, culture media and FectoPRO (Polyplus) would be added to cells to a ratio of 0.5 ...
Four liters of bacterial culture yielded 1 ml of 39 μM RAD52. Protein labeling For fluorescence labeling of trimeric RPA complexes, the purified protein complexes were incubated with Sulfo-Cyanine3 maleimide (Luminoprobe) using an excess molar ratio of 40:1 (label:protein) overnight at 4...
Springtail-inspired omniphobic slippery membrane with nano-concave re-entrant structures for membrane distillation Here, authors successfully fabricated a springtail-inspired nano-concave reentrant structured polystyrene (PS) membrane using electrospraying technology followed by lubricant dip-coating, which endowe...
In brief, the pellet from every one-liter fresh cell culture was deciliated with 3 mM dibucaine (Sigma-Aldrich) in 150 ml fresh SSP medium and centrifuged at 2,000g for 10 min to remove the cell body. The cilia were spun down from the supernatant at 12,000g for 10 min, ...
Protein backbone atoms were constrained using a spring force (in form of k(∆x)2)83 and movement of hydrogen atoms were constrained by SHAKE algorithm86. The systems were then simulated for 5 ns in a NPT ensemble with a pressure of 1 bar applied using Berendsen barostat87 and ...
Uniformly 15N-labeled (or 13C- or 15N-labeled) protein samples were prepared in M9/H2O medium supplemented with 1 g liter−1 of 15NH4Cl and 2 g/L of glucose (or 13C-glucose). 2H-Methyl-13CH3-Met-labeled samples were prepared according to published methods35,36. The cells were ...
One milliliter of culture was fixed with 2.5% formaldehyde and visualized at the fluorescence microscope (Leica DC 350F). More than 200 cells were analyzed in each experiment. Reporting Summary Further information on research design is available in the Nature Research Reporting Summary linked to ...
The real CV is tethered to its fictitious CV via a stiff spring with a large force constant and the adaptive biasing force is equal to the running average of the negative of the spring force. The biasing force is only applied to the fictitious CV, which in turn "drags” the real ...
Engineered yeasts producing squalene or mogrol were incubated overnight in 3 mL YPD medium at 30 °C with shaking at 250 rpm as seed cultures. One milliliter seed culture was used to inoculate 50 mL of YPD medium with or without 30 μg mL−1terbinafine (Aladdin, China) fo...
Droplet impact on solid surfaces is essential for natural and industrial processes. Particularly, controlling the instability after droplet impact, and avoiding the satellite drops generation, have aroused great interest for its significance in inkjet pr