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A pure F1 diploid colony from each pair of diploid parents was picked from the YEPD plate, propagated clonally in YEPD and frozen for later use.
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Resistant colonies were picked onto mouse embryonic fibroblast-coated 24-well plates, propagated and analyzed for mCh expression.
Single dauer worms were plated onto small NGM plates, propagated for approximately two weeks, and verified for GFP expression using a Leica DMI3000B.
P-to-S mode-converted waves at the upper surface of the plate propagate through the slab crust for longer distances than do direct P- and S-waves; therefore, their arrival-time data are convenient for estimating the seismic velocity structure within the slab crust.
On day 8, cells from duplicate wells were restimulated separately with 1×106 allogenic, irradiated PBMC, 10 U/mL IL-2, 1 ng/mL IL-15, and 1 µg/mL phytohemagglutinin (Remel) in a 48-well plate and propagated as four separate T-cell lines.
Resistant colonies were collected and transferred to a fresh plate and propagated.
One colony from the MRS agar plate was propagated into MRS broth and incubated at 37°C for 24 h.
All 2° sublines and 1° pseudolines were replicated 5X from immature individuals taken directly from the thawed plate and propagated for three generations by single-individual descent (P1-3) at 4- or 5-day intervals, respectively.
A single colony of S. aureus was selected from the agar plate and propagated in LB overnight to reach an optical density of 1.11 at 578 nm which corresponds to 2 × 10 CFUs/mL.
Control, SPLhi or SPLlo cells 0.5 × 10 were plated in six-well plates and propagated for 24 h.
Clones at generation approximately 50, 100, and 200 were plated from frozen samples onto sulfate-limiting plates and propagated in sulfate-limiting liquid medium to attempt to maintain selection for the amplicons that arose during chemostat growth.
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