Our data suggests that for species unrelated to Arabidopsis it would be better to use both introns predicted and empirical data for assay design to maximize validation rate and evaluate genetic diversity.
The purpose here is to develop the theory of lethal mutagenesis as it applies to the types of empirical systems and assays that may be easily employed with viruses, with the specific goal of resolving the question.
Visual observations in the field did not distinguish mutant 2G from the susceptible Thatcher parent, however, a 30% reduction in chitin accumulation was observed which likely reflects the sensitivity of the chitin assay over empirical visual observations.
The 'Luria Delbrück' distribution describing the number of mutants appearing in a growing population before screening is widely applied in estimating mutation rates from fluctuation assays (see Empirical approaches), including long-standing consideration of drug resistance mutations (David 1970).
This study therefore highlights the importance of selecting a stable reference gene for normalization of target gene expression based on empirical evaluation for a given assay condition, cell type and infection status.
The biomarker-strategy design, in which patients are randomized to two arms (assay-directed vs empirical treatment), is a reasonable approach for evaluating multiple markers and has been recommended for evaluation of chemoresponse assays (Blue Cross and Blue Shield Association, 1995, 2000, 2002; Schrag et al, 2004; Burstein et al, 2011).
In the third example, Huttenhain et al. [ 18] extended this empirical testing to develop MRM assays for over 1,000 cancer-associated proteins in both serum and urine.
However, the design of these assays remained mostly empirical, being notably based on the use of the 16S rRNA gene [ 3], until bacterial genome sequencing became a reality in the mid-1990s [ 4].
Future investigations conducted prospectively to analyze the diagnostic accuracy and utility of these assays should be conducted before empirical antimicrobial treatment is replaced by specific treatment targeting organisms identified in molecular assays.
