TABLE 1.
Parameter | Assay value for indicated gene
|
|||
---|---|---|---|---|
ltrBE1 | ltrB | ltrA | oppD | |
PCR efficiency determined by usingb: | ||||
Recombinant plasmid DNA | 1.97 | 1.96 | 1.91 | 1.96 |
cDNA templates | 1.92 | 1.95 | 1.98 | 1.94 |
Melting temp (°C) | 78.5 | 79.0 | 77.0 | 78.0 |
Variationc (%) | ||||
Intra-assay | 1.7 | 1.3 | 1.6 | 3.9 |
Interassay | 3.0 | 3.9 | 2.7 | 4.2 |
PCR efficiency (E) is calculated according to the following equation: E = 10(−1/slope). The maximal efficiency of PCR is 2 where every PCR product is replicated every cycle.
PCR efficiency was determined by using recombinant plasmid DNA containing target genes as templates and by using cDNA templates generated from in vitro-transcribed total L. lactis ML3 RNA.
Intra-assay variations were determined in three repeats within one real-time PCR run. Interassay variations were determined in four different experiment runs in 4 days. Intra-assay and interassay variations were determined by using recombinant plasmid DNA containing target genes as templates.