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The same volume of cell extract/sample was used for all experiments as well as for SDS-PAGE.
Briefly, 105 to 107 cells/25 µl per well of a 96 well plate were lysed in the same volume of cell lysis reagent by incubation for 5 min at RT. Fifty microlitres of the reconstituted luciferase reagent was added to the well by automated injection (BD Bioscience, Monolight luminometer).
These data were used to make signal-to-input profiles (as in Fig. S2), i.e., a kind of comparative genomic hybridization (CGH) comparing immunoprecipitated DNA to input DNA from the same volume of cell lysate.
The base procedure was performed in two models: (1) measuring total emitted fluorescence by staining same volume of cell suspension for different treatments (cells at the lag stationary phase), which would quantify total produced lipid volumetrically; (2) measuring emitted fluorescence by staining same cell number (10 cells) for different treatment, which would quantify stored lipid in cells.
At this stage, dogs were randomized (in a 2 1 ratio of olfactory mucosal cell: medium alone) to receive either olfactory mucosal cell in cell transplant medium or the same volume of cell transport medium alone, injected directly into the parenchyma of the spinal cord.
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A total of 1.5 × 10 cells in 10 μL of EMEM media or the same volume of cell-free PBS (control) were injected in the dorsal prostate lobe using a 30 G needle.
For fluorescent assay of the three-input biosensor comprising two cell consortia, the two cell populations were diluted from overnight culture to the same cell density at OD600=0.025 and mixed using the same volume of each cell culture prior to being loaded into microtitre plate wells.
Result of total lipid measurement using microplate fluorometryin the same volume of the cell suspension was summarized in Figure 4.
The reagent method resulted in isolation of somewhat more exosomes from the same volume of HeLa cell culture media, from which ~7.5 ng RNA was extracted.
The same volume of the cell lysis reagent was added; further, the lysates were incubated at room temperature for 5 min.
Concerning neuroblasts, mutants and controls displayed the same volume of migrating cells at 4 months.
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