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According to the model calculation, the original mixture contained approximately 60% seawater and its volume subsequently was reduced through evaporation by around 30%.
The 20-μl mixture contained approximately 50 ng of total DNA, 5 mM each of dNTPs, 20 pmol each of forward and reverse primers, and 0.5 U of Taq DNA polymerase.
The 20-μl PCR mixture contained approximately 50 ng of total DNA, 5 mM each of dNTPs, 20 pmol of each forward primer and reverse primer, and 0.5 U of Taq DNA polymerase.
The PCR reaction mixture contained approximately 100 ng of DNA, 1× PCR buffer, 1.5 mM MgCl2, 200 µM of each dNTP, 0.2 µM of each primer, and 1 U of Platinum Taq DNA polymerase (Invitrogen, Carlsbad, CA) in a final volume of 50 µl.
Each reaction mixture contained approximately 10 ng of template DNA; 0.4 pmol (each) forward (ITS1) and reverse (ITS4) primers; 10 μM (each) dATP, dCTP, dGTP, and dTTP; 10X reaction buffer containing 1.5 mM MgCl2 (AB) and 2 U of Taq Gold (AB).
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It is found that the explosion length scaling of Ro ∼ 26λ becomes also invalid when the mixture contains approximately this same amount of argon dilution or more.
For competition experiments, a mixture containing approximately 3×107 CFUs of wild type parent and 3×107 CFUs of isogenic Δpvl mutant were used to co-infect New Zealand white rabbits via the marginal ear vein.
To initiate each of the 12 sub-lineages, a 50 μl aliquot of the founding population mixture, containing approximately 5 million conidia, was inoculated into the end of glass tubes (2.5 × 30 cm) filled horizontally with 20 ml of solid media (1.5% agar).
PCR was performed in a 25 μl reaction mixture containing approximately 100 ng DNA, 1.0 μM each primer, 0.2 mM each dNTP, 2.0 mM MgCl2, 1.0 U Taq DNA polymerase with 1 × reaction buffer (Promega, Madison, WI), and 2% dimethyl sulfoxide.
The DOPG EYPC lipid mix contains 66% 18∶1 acyl chains, while the PBPS EYPC and EYPC mixtures contain approximately 33%.
PCR amplification was performed in 50-µl reaction mixtures containing approximately 50 75 ng genomic DNA templates, 1.5 mM MgCl2, 0.2 mM of each dNTP, 1 µM of each primer, 0.1 mg BSA/ml and 1 unit Taq DNA polymerase.
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