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In LBX medium the colonies looked domed and were picked for digesting to derive cell lines.
In KOSR medium the colonies looked flattened and were picked for cutting to derive cell lines.
In LBX medium, the small colonies formed as early as the fifth day after induction, whereas, in KOSR medium, the colonies were firstly visualized at the eighth day after induction (Fig. 1B).
After removing the small compounds from LBX medium, the colonies had a tendency to become flattened (Wang et al., 2013), suggesting that these small molecular compounds are responsible for mouse ESCs-like colony formation and cell proliferation.
After 15 days of incubation in complete culture medium, the colonies were stained with crystal violet after fixation with formaldehyde and were counted manually.
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The yeast strain YIGLO3GCS1 was constructed by the following steps: The amdSYM marker was removed from YIGLO3 genome in YPD medium, and the colonies were counter-selected on a fluoroacetamide plate as described (Solis-Escalante et al. 2013).
The intracellular survival was determined by plating serially diluted cultures on 7H10 medium and the colonies were enumerated after 3 days.
At each passage, the cultures were diluted and plated on AS168 nonselective medium, and the colonies were subsequently patched on mevinolin-selective plates to determine the fraction of mevinolin-resistant cells.
A change in the color of the medium surrounding the colony, from red to yellow, indicated arabinose fermentation.
After 24 h of growth in SD -trp- leu (DDO, double dropout medium) at 30°C, the colonies were subsequently plated onto SD-trp-leu-ade-his medium (QDOdruple drop-out (QDO) medium).
The fraction of cells retaining the plasmid pMA5-RDPE after 80 generations in nonselective medium (SR without kanamycin) was 0%, whereas in selective medium 85% of the colonies still contained the plasmid.
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