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ChIP material was washed three times in 100 µl RIPA and once in 100 µl TE buffer, and transferred into a new tube while in TE.
Cell pellets were washed with TE buffer, resuspended in TE buffer and transferred into screw top tubes containing 500 μl of glass beads.
Ten microliters of this ligation mix were premixed with 30 μl PCR buffer and transferred into a PCR machine at 60°C.
Sample was diluted 1 1 with Binding buffer and transferred into large-volume sample loop (Superloop 50 mL, GE) for sample loading.
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After 30 45 min incubation on ice, the cells were washed twice with MACS-buffer and resuspended at 1×106 cells per 0.5 ml MACS-buffer and transferred into a 4 ml tube.
After centrifugation at 3,500× g for 2 min, the final worm pellet was resuspended in the same lysis buffer (1/1.5 of V worms/V buffer ratio) and transferred into eppendorf tubes.
The silica disks supporting the DC-SIGNR and the 5 mol % Man9GlcNAc2-DPPE bilayers were maintained under buffer A and transferred into the chamber surface force apparatus, which was filled with buffer A. All measurements were performed at 21 ± 0.2 °C.
Of this 500 µl sample 200 µl was transferred into a HPLC reaction vial and diluted 1 1 with 200 µl 0.05 M pH 6.8 phosphate buffer solution and transferred into the HPLC machine for analysis.
The biosensors were then washed in Buffer A and transferred into 96-well plates (Greiner #655209) containing a 7-point, 1.5-fold serial dilution of the TRIM24 protein (starting from a 67 nM solution in Buffer A).
The lipid droplet-containing supernatant was adjusted to 0.33 M sucrose in a final volume of 3 ml, using a 1 M sucrose solution in buffer A, and transferred into a 12 ml ultracentrifugation tube.
Both tissues were washed with phosphate buffered saline and transferred into a Petri dish.
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