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Ten grams from each dry, powdered sample were used for protein digestion with 1 g of papain (Merck, Darmstadt, Germany) in 400 mL 0.1 M sodium acetate (pH 6.0) with 5 mM ethylenediamine tetra-acetic acid, and 5 mM cysteine.
Two grams from each tissue were washed with isotonic saline solution and homogenized (Janke & Kunkel Ultra Turrax T25) in 5 ml saline.
Five grams from each callus were mixed with 10 ml extraction buffer [100 mM N-2-hydroxyethylpiperazine- N-2-ethanesulfonic acid (HEPES -KOH (pHEPES -KOHmM ethylenediaminetetraacetic acid (EDTA), and 0.05% (v/v) Triton-X-100] and homogenized for 20 s using a Waring blender.
According to the FDA [ 30], 50 grams from each sample was placed in ethylene oxide gas-sterilized polypropylene bags (Whirl-Pak, Nasco, USA) and 450 mL of lactose broth was added in order to achieve a final dilution 1 : 10 (10−1).
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Approximately 1 gram (g) from each sample was used for bacterial cultures.
0.05 gram from each feed stock was added to 5 g of deionized water to make a 1 wt% mixture.
For measuring Zn residues in the investigated fish organs, a gram from each organ was placed in crucible and ashed in a muffle furnace (Thermolyne Corporation, Dubuque, Iowa, USA) for 6 h.
For model building our tool considers either the entire set (100%) or a partial (75%, 50%and25%5%) set of non-repeatable n-grams from each genome.
For testing both models, we used 1% of the total n-grams from each bacterial genome, which also contain plasmid sequences as expected in the true metagenomic samples.
The n-grams from each genome were compared against the n-grams in the other genomes to finally arrive at a set of unique (present in a single genome) and common (present in multiple genomes) n-grams.
One gram from each sample was suspended in 9 ml sterile water (of 0.9%, 10%, and 20% NaCl w/v) and serial dilutions to 10−4.
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CEO of Professional Science Editing for Scientists @ prosciediting.com