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To the Editor: Thanks to Frederick Turner ("Autumn Is Mellow Deep in Normandy," Sept. 10) for reminding me of a visit, in 1976, to the splendid Calvados countryside and an overnight at the same Auberge du Vieux Puits in Pont-Audemer.
The strain was aseptically stored in fluid thioglycollate medium overnight at the same environment.
After electrophoresis on an agarose gel, RNA was transferred to a nylon membrane followed by a UV crosslink for 10 min. After prehybridizing for 2 h at 68°C, blots were hybridized with the digoxigenin-labeled probe overnight at the same temperature.
After 2 h at 60°C either the fluoresceine-labelled LplA coding region, the human DHFR (hDHFR) or the blasticidin S deaminase (BSD) probes, generated using the Gene Images Random Prime labelling module according to the manufacturer's recommendation (GE Healthcare), were added and the blots hybridised overnight at the same temperature.
Then, the P-labeled probe was added and hybridization continued overnight at the same conditions.
Probe hybridization with DIG-labeled probes (200 ng/ml) was done overnight at the same temperature.
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A force powerful enough to change landscapes and age-old cultures overnight, but at the same time so fragile, tourism is so totally dependent on image, that even a perception of discomfort -- not to mention a real disaster -- can damage it.
Cells were washed and infected with S. Typhimurium in the presence of the yeast (overnight or at the same time of bacterial infection) or not.
Staffing levels overnight were maintained at the same ratios as if patients were dialysing during the day.
In parallel, bacteria from the control plates were collected and incubated on LB and LB agar at 37°C overnight in the same incubator.
The slides were incubated at 4°C overnight in the same buffer containing an alkaline phosphatase-conjugated anti-digoxigenin antibody (Roche) diluted at 1 1000.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com