Aims: The aim of this study was to evaluate the variability in lots of a DNA extraction kit using real‐time PCR assays for Bacillus anthracis, Francisella tularensis and Vibrio cholerae.
Methods and Results: Replicate aliquots of three bacteria were processed in duplicate with three different lots of a commercial DNA extraction kit. This experiment was repeated in triplicate. Results showed that cycle threshold values were statistically different among the different lots.
Conclusions: Differences in DNA extraction reagent lots were found to be a significant source of variability for qPCR results. Steps should be taken to ensure the quality and consistency of reagents. Minimally, we propose that standard curves should be constructed for each new lot of extraction reagents, so that lot‐to‐lot variation is accounted for in data interpretation.
Significance and Impact of the Study: This study highlights the importance of evaluating variability in DNA extraction procedures, especially when different reagent lots are used. Consideration of this variability in data interpretation should be an integral part of studies investigating environmental samples with unknown concentrations of organisms.
Sediment-Record Calculations (SEDCALC), a menu-driven set of interactive computer programs, was developed to facilitate computation of suspended-sediment records. The programs comprising SEDCALC were developed independently in several District offices of the U.S. Geological Survey (USGS) to minimize the intensive labor associated with various aspects of sediment-record computations. SEDCALC operates on suspended-sediment-concentration data stored in American Standard Code for Information Interchange (ASCII) files in a predefined card-image format. Program options within SEDCALC can be used to assist in creating and editing the card-image files, as well as to reformat card-image files to and from formats used by the USGS Water-Quality System. SEDCALC provides options for creating card-image files containing time series of equalinterval suspended-sediment concentrations from 1. digitized suspended-sediment-concentration traces, 2. linear interpolation between log-transformed instantaneous suspended-sediment-concentration data stored at unequal time intervals, and 3. nonlinear interpolation between log-transformed instantaneous suspended-sediment-concentration data stored at unequal time intervals. Suspended-sediment discharge can be computed from the streamflow and suspended-sediment-concentration data or by application of transport relations derived by regressing log-transformed instantaneous streamflows on log-transformed instantaneous suspended-sediment concentrations or discharges. The computed suspended-sediment discharge data are stored in card-image files that can be either directly imported to the USGS Automated Data Processing System or used to generate plots by means of other SEDCALC options.
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