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All colonies from each plate, or within a defined section, were picked in an unbiased manner for DNA extraction and 16S rRNA gene sequencing using the universal primers: 7F, 5′-AGAGTTTGATYMTGGCTCAG-3′; 926R, 5′-ACTCCTACGGGAGGCAGCAG-3′.
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The ribbons with samples were placed into a water bath (40 °C) and the best section was picked up by glass slides and placed on a slide drier (20 °C) for overnight.
Each section was picked up with a chrome alum gelatin-coated slide.
One block in each census section was picked by lot; two women were interviewed in each block after the randomization method described previously [ 20, 21].
Sections were picked onto Superfrost Plus or Apes coated Superfrost slides.
Ultrathin plastic sections were picked up on copper grids and contrasted with uranyl acetate and lead citrate.
After cryo-sectioning, sections were picked up and thawed according to Liou et al. [28] in a 1∶1 mixture of 2.3M sucrose and 2% (v/v) methylcellulose.
Sections were picked up from the knife with a loop dipped in 2.3 M sucrose and transferred to a formvar/carbon coated copper grid.
Sections were picked up on copper grids and stained in 2% uranyl acetate (12 min) and Reynolds' lead citrate (6 min).
Sections were picked onto 200 mesh copper grids, stained with uranyl acetate and lead citrate and examined under a Tecnai Twin (FEI) electron microscope.
Ultracryomicrotomy was carried out at −100°C on a Leica Ultracut UCT with EM FCS cryoattachment (Leica, Bannockburn, IL) using a Diatome diamond knife (Diatome US, Fort Washington, PA). 60 70 nm frozen sections were picked up with a 1∶1 mixture of 2.3 M sucrose and 2% methyl cellulose and transferred onto Formvar and carbon-coated copper grids.
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