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. 2025 May 9;15:16184. doi: 10.1038/s41598-025-98444-8

Table 4.

Feature descriptions.

Features represented within mutated templates
Feature description Feature abbreviation Notes
Percent of substitutions in the first primer % MM P1 If a template only has mismatches in a single primer, that primer is labeled as first primer. If both primers contain mismatches, then primers will be randomly labeled as first primer and the other second primer
Percent of substitutions in the second primer % MM P2
Percent of substitutions in the probe % MM Probe
Percent of deletion in the first primer # Del P1
Percent of deletion in the second primer # Del P2
Percent of deletion in the probe # Del Probe
Nucleotide distance from the 3′ end of the first primer to the nearest substitution/deletion Nearest 3′ P1
Nucleotide distance from the 3′ end of the second primer to the nearest substitution/deletion Nearest 3′ P2
Number of deletion and substitution within 5 nucleotide distance to the 3′ of the first primer # MM/Del 3′ P1
Number of deletion and substitution within 5 nucleotide distance to the 3′ of the second primer # MM/Del 3′ P2
Difference in annealing temperature of the first primer and protocol annealing temperature (55 °C) Temp Diff P1 Annealing temperatures were calculated using the Tm_GC function of the biopython package35
Difference in annealing temperature of the second primer and protocol annealing temperature (55 °C) Temp Diff P2
Difference in annealing temperature of probe and protocol annealing temperature (55 °C) Temp Diff Probe