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HSP_NumtS not present in the European sample were validated in an Ethiopian sample whose mtDNA belonged to the L0 haplogroup.
Field and museum identifications for each sample were validated by amplifying and sequencing a portion of the mitochondrial cytochrome b (Cytb) gene using the methods of Stadelmann et al. (2007).
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The concentration of each sample was validated using the Nanodrop spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA).
The enriched target DNA in each library sample was validated and quantified by microfluidics analysis using the Bioanalyzer High Sensitivity DNA Assay kit (Agilent Technologies) and the 2100 Bioanalyzer with the 2100 Expert Software.
Samples were validated in lung tissue achieving a validation rate of 65.7% (Supplemental Table S3).
The eDNA samples were validated by endpoint PCR using the best molecular marker for dreissenid specificity previously selected in Stage A. Gradient PCR with an annealing temperature ranging from 60 to 70 °C and a Touch-Down PCR with decreasing annealing temperature from 70 to 60 °C (−1 °C/cycle in the first ten cycles) were performed in parallel to optimize PCR amplification.
All samples were validated for subsequent analysis with a RIN value of at least 6.0.
Next, bearing area and pullout strength relationships developed from the test samples were validated against nine commercially available suture anchors, including the Mitek QuickAnchor and SpiraLok, Opus Magnum(2), ArthroCare ParaSorb, and Arthrex BioCorkscrew.
All the samples were validated in duplicates.
Results from clinical samples were validated using PCR.
Results of selected samples were validated using the Epitect Sequencing Service (Qiagen).
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