Table 3.

Effect of primer degeneracies on PCR product ratiosa

PrepnEco(GC)/Eco(AT)m ratiob
15 cycles25 cycles35 cycles
Template mixture1.0 ± 0.041.0 ± 0.041.0 ± 0.04
PCR 11.4 ± 0.161.5 ± 0.062.3 ± 0.10
PCR 21.4 ± 0.141.8 ± 0.112.3 ± 0.07
PCR 31.3 ± 0.111.7 ± 0.072.5 ± 0.20
PCR 41.4 ± 0.261.9 ± 0.071.9 ± 0.14
PCR 51.3 ± 0.161.8 ± 0.092.0 ± 0.17
  • a The data were generated by dot blot hybridization by using the template-specific probes Eco and EcoM and mixtures containing equal amounts of templates Eco(GC) and Eco(AT)m. PCR products were amplified from the same template mixtures for 15, 25, and 35 cycles.

  • b Means ± standard deviations were calculated from data for three replicate dots for each of the samples which was hybridized with the specific probes and quantified by phosphorimaging.