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The protein content of MV preparations was quantified by Bradford method (Bio-Rad, Hercules, CA, USA).
The total amount of protein in extract preparations was quantified using the Bradford protein assay (Bio-Rad).
Protein concentration in the MAb preparations was quantified using Coumassie Plus Protein Reagent, murine IgG concentration was determined using an Antibody Assay kit (both obtained from Pierce Biotechnology, Rockford, IL).
FMDV RNA, within these RNA preparations, was quantified using two different real time RT-PCR assays, that target either the 5' untranslated region or the 3D coding region, as described previously [8], [9], [10].
The protein content of EV preparations was quantified by Quantifluor (Promega) using NanoOrange Protein Quantitation Kit (Life Technologies, Carlsbad, CA).
Nucleosomal and ChIP DNA from independent preparations was quantified by Real Time quantitative PCR (ABI Prism) for validation of nucleosome shifts in different conditions.
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Preparations were quantified and their purity was determined by standard spectrophometric methods.
DNA preparations were quantified using PicoGreen reagents according to the supplier instructions.
RNA preparations were quantified by absorbance at 260 nm (A260) using a Nanodrop spectrophotometer (Labtech Intl., E. Sussex, UK).
The amounts of HA and NA proteins into purified virion preparations were quantified by western blot analysis as an estimate of their relative amounts of incorporation into HIV pseudoparticles.
The final pellet of product RNA was resuspended using 100 µl of RNase free water (Ambion, Austin, TX) and incubated at 65°C for 5 min. Bacterial genomic DNA preparations were quantified using NanoDrop ND1000 (Thermo Scientific Inc., Waltham, MA) spectrophotometer and genome copy number was calculated using the genome size and the DNA concentration.
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