TABLE 1.

Quantities of chromosomal and plasmid DNA used to construct qPCR standard curves

Target DNAAbbreviationLengthaTested concn range (ng/μl)bNo. of target gene copies tested (range [per μl DNA])c
Dehalococcoides sp. strain BAV1
    Genomic DNABAV1 DNA1.5 Mb2 × 10−6-201.2 × 100-1.2 × 107
    Cloned 16S rRNA genepBAV1/16S5.4 kb1 × 10−8-11.4 × 100-1.4 × 108
    Cloned bvcA genepBAV1/bvcA5.0 kb1 × 10−8-11.7 × 10−1-1.7 × 108
Dehalococcoides sp. strain GT
    Genomic DNAGT DNA1.5 Mb1.2 × 10−6-127.3 × 100-7.3 × 106
    Cloned vcrA genepGT/vcrA5.4 kb1 × 10−8-11.8 × 100-1.8 × 108
Dehalococcoides sp. strain FL2
    Genomic DNAFL2 DNA1.5 Mb5 × 10−6-503.0 × 100-3.0 × 107
    Cloned 16S rRNA genepFL2/16S5.4 kb1 × 10−8-11.7 × 100-1.7 × 108
    Cloned tceA genepFL2/tceA7.3 kb1 × 10−8-11.25 × 100-1.25 × 108
Dehalococcoides ethenogenes strain 195 genomic DNA195 DNA1.5 Mb6 × 10−6-603.6 × 100-3.6 × 108
Anaeromyxobacter dehalogenans strain 2CP-C genomic DNAd2CP-C DNA5.0 Mb2.4 × 10−6-24.58.9 × 100-8.9 × 106
  • a Based on the genome sizes of Dehalococcoides ethenogenes strain 195 (1.5 Mb) and Anaeromyxobacter dehalogenans strain 2CP-C (5.0 Mb) or calculated by adding the size of plasmid pCR2.1 (3.9 kb) to the cloned insert size (1.5 kb for the BAV1 16S rRNA and bvcA genes and 3.4 kb for the tceA gene).

  • b Based on spectrophotometric quantification of the DNA at 260 nm and 10-fold serial dilutions.

  • c Calculated by equation 1 as described in the text.

  • d Two 16S rRNA genes per genome.