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The colony grown on SD selection medium were picked up and then confirmed by the PCR method using the primer pair up100-F and hstg2-R whether the expression plasmid was inserted into the genomic DNA (Table 1).
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For this purpose, a patch of colony on agar medium was picked to culture in liquid medium for three days; different stress conditions were then applied by adding sorbitol or NaCl, or by transfer to liquid medium lacking glucose or nitrate, and the cells were cultured for an additional eight hours.
After 4 days on selective medium, segregants were picked and grown 2 days in YPD in a 96-well plate for sequencing and frozen stocks.
Ascospores from Dp EB4) a × E A were germinated on adenine-supplemented Vogel's FGS medium, 130 germlings were picked to adenine-supplemented Vogel's glucose medium, and then their growth was tested on unsupplemented Vogel's glucose medium.
Measurements of cell size and DNA content were of cells that had been selected on 5-FOA plates, then transferred to YPD medium before clones were picked to inoculate the initial growth cultures examined.
After immobilization on 2.5% agarose pads, medium-budded cells were picked and single stack images were acquired.
Clonality was obtained on semi-solid agar plates: cells from the original strains were cultured in semi-solid agar K medium and individual colonies were picked off and transferred into new semi-solid agar medium.
Peaks with high, medium or low heights were picked for validation.
Colonies were picked into liquid medium with TM, grown at 51°C, and plated in medium supplemented with TM and FUdR.
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.
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