Table 1.
Determination of error rate values for B35DNAP and B35-HhH during MDA of plasmid DNA in the presence of Mg2+ (A) or Mg2+/Mn2+ (B) ions.
| Sample | Experiment | Colonies | f | d | Error rate (× 10–6) | |
|---|---|---|---|---|---|---|
| A. Cofactor: MgCl2 10 mM | ||||||
| Control | 1 | 9,894 | 0.18% | |||
| 2 | 15,480 | 0.08% | ||||
| 3 | 8,496 | 0.56% | ||||
| 4 | 6,980 | 0.62% | ||||
| 5 | 1608 | 0.56% | ||||
| B35DNAP | 1 | 6,598 | 2.88% | 11.88 | 6.64 | |
| 2 | 10,248 | 2.84% | 11.54 | 7.00 | ||
| 3 | 5,380 | 4.63% | 11.97 | 9.93 | ||
| 4 | 3,010 | 6.18% | 12.14 | 13.40 | ||
| 5 | 4,176 | 6.66% | 10.97 | 16.26 | ||
| B35-HhH | 1 | 17,441 | 0.72% | 14.05 | 1.13 | |
| 2 | 13,511 | 0.68% | 13.84 | 1.27 | ||
| 3 | 6,065 | 0.99% | 13.91 | 0.89 | ||
| 4 | 3,631 | 2.26% | 13.42 | 3.58 | ||
| 5 | 9,462 | 3.89% | 14.02 | 6.94 | ||
| Average | B35DNAP | 4.64% | 11.70 | 10.64 ± 3.71 | ||
| B35-HhH | 1.71% | 13.85 | 2.76 ± 2.30 | |||
| B. Cofactor: MgCl2 10 mM + MnCl2 10 nM | ||||||
| Control | 4 | 6,980 | 0.62% | |||
| 5 | 1608 | 0.56% | ||||
| B35DNAP | 4 | 2,938 | 6.06% | 12.27 | 12.97 | |
| 5 | 3,868 | 5.95% | 11.03 | 14.27 | ||
| B35-HhH | 4 | 3,450 | 1.19% | 14.08 | 1.19 | |
| 5 | 4,551 | 3.69% | 14.02 | 6.53 | ||
| Average | B35DNAP | 4.00% | 7.77 | 13.62 ± 0.65 | ||
| B35-HhH | 1.63% | 9.36 | 3.86 ± 2.67 | |||
Mutant frequencies (f) were determined by dividing the total number of white plaques by the total number of plaques. Template doublings (d) were calculated using the equation d = log2((ng product)/(ng input)). Background frequency (f0) was calculated in control experiments with plasmid DNA purified from bacteria, digested and re-ligated in the same conditions (“Materials and methods”). Error rate was calculated using the equation ER = (f − f0)/bp·d, where bp is the number of detectable sites in the lacZ gene (342 bp).