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Candidate targets of interest were verified by qPCR using SYBR green detection.
The events of interest were verified by death certificates and medical records, therefore misclassification is unlikely.
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Expression of the protein of interest was verified by Western blot.
The presence of the fragment of interest was verified by RsaI and EcoRI restriction digestion.
Normal distribution of the variables of interest was verified with the Kolmogorov-Smirnov test.
Enrichment for cells of interest was verified by microscopic examination of the LCM cap after microdissection.
The presence of the proteins of interest was verified by specific secondary antibodies conjugated to horseradish peroxidase (HRP).
Plasmid DNA was prepared using the Qiagen Miniprep Kit (Qiagen), and the presence of the fragment of interest was verified by NOT1 restriction enzyme digestion.
Successful and specific amplification of the region of interest was verified by visualising 5 μl of the PCR product on a 2% agarose gel.
The representativeness of microarray probe hybridization and specific gene expression responses of toxicological interests were verified using qPCR (N = 6).
In brief, replicate PCRs were performed amplifying 100 bp to 200 bp around the coding SNP of interest; products were verified by agarose gel electrophoresis and sequenced using ABI Big Dye chemistry and capillary electrophoresis on an ABI 3730 sequencer (Applied Biosystems).
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CEO of Professional Science Editing for Scientists @ prosciediting.com