Error Rate Comparison during Polymerase Chain Reaction by DNA Polymerase

Table 2

Error rate values for six PCR enzymes included in this study are presented. Each enzyme was used in two independent PCR reactions. Average doublings/PCR reaction () is the average of doubling values for each of the 94 PCR reactions in one plate, where doublings are calculated from the formula = (ng DNA after PCR/ng DNA input). Error rate (f) is calculated as f = / (target size ×  ), where is the number of mutations observed for all clones that were sequenced and the (target size ×   for each target that was cloned.

Enzyme	Expt.	Avg. doublings/PCR reaction	Number of clones sequenced	Total bp sequenced	Number of mutations observed	Error rate
Taq	1	20.5 ± 1.2	65	8.8 × 10^4	54	3.0 × 10^−5
	2	16.7 ± 0.7	37	4.7 × 10^4	45	5.6 × 10^−5
AccuPrime-Taq	1	17.0 ± 1.2	75	1.0 × 10^5	18	1.0 × 10^−5
	2	16.9 ± 0.6	N.D.	N.D.	N.D.	N.D.
KOD	1	20.8 ± 1.5	70	1.0 × 10^5	16	7.6 × 10^−6
	2	17.6 ± 0.8	N.D.	N.D.	N.D.	N.D.
Pfu (cloned)	1	16.5 ± 1.1	151	2.0 × 10^5	9	2.8 × 10^−6
	2	12.0 ± 1.8	N.D.	N.D.	N.D.	N.D.
Phusion	1	21.0 ± 1.9	175	2.4 × 10^5	13	2.6 × 10^−6
	2	16.6 ± 1.1	N.D.	N.D.	N.D.	N.D.
Pwo	1	22.5 ± 1.2	170	2.4 × 10^5	13	2.4 × 10^−6
	2	17.6 ± 0.6	N.D.	N.D.	N.D.	N.D.

N.D.: not determined.