Table 1.
Error rate estimates from two independent experiments. The table shows the number of mismatches that remain after consensus sequence assembly for reads tagged with the same UMI, the total number of unique UMI tags observed, and error rate estimates for 20 (25 for Phusion) cycles of PCR amplification with a 150-bp template. Error rate estimates are provided as the number of erroneous bases per template base per cycle, with 95% confidence intervals (CI) calculated using normal approximation for binomial proportions. Error rateLA represents estimates for the linear amplification step, while Error ratecorr represents the estimate of conventional PCR error rate after correcting for linear amplification errors.
| Polymerase* | Exp. | Error count | UMI tags | Error rate, x10 −5 [95% CI] | Error rate LA , x10 −5 | Error rate corr , x10 −5 [95% CI] |
|---|---|---|---|---|---|---|
| Encyclo | 1 | 24557 | 185560 | 4.30 [4.25, 4.35] | 12.79 | 3.68 [3.63, 3.73] |
| Encyclo | 2 | 14211 | 101516 | 4.55 [4.48, 4.62] | 3.93 [3.86, 3.99] | |
| Kapa HF | 1 | 339 | 7876 | 1.40 [1.25, 1.55] | 8.17 | 1.00 [0.88, 1.13] |
| Kapa HF | 2 | 2519 | 57052 | 1.44 [1.38, 1.49] | 1.04 [0.99, 1.08] | |
| Phusion | 1 | 30 | 1348 | 0.72 [0.47, 0.98] | 4.99 | 0.48 [0.27, 0.69] |
| Phusion | 2 | 23 | 1351 | 0.55 [0.33, 0.78] | 0.31 [0.14, 0.48] | |
| SD-HS | 1 | 6714 | 33076 | 6.60 [6.46, 6.74] | 26.89 | 5.29 [5.16, 5.42] |
| SD-HS | 2 | 10362 | 58518 | 5.76 [5.66, 5.86] | 4.45 [4.36, 4.54] | |
| SNP-detect | 1 | 457 | 13870 | 1.07 [0.97, 1.17] | 5.04 | 0.83 [0.74, 0.91] |
| SNP-detect | 2 | 848 | 32310 | 0.85 [0.80, 0.91] | 0.61 [0.56, 0.66] | |
| Taq-HS | 1 | 1875 | 15137 | 4.03 [3.86, 4.20] | 18.36 | 3.13 [2.98, 3.29] |
| Taq-HS | 2 | 3113 | 24082 | 4.20 [4.07, 4.34] | 3.31 [3.18, 3.43] | |
| Tersus-buffer1 | 1 | 2550 | 46927 | 1.77 [1.70, 1.83] | 5.98 | 1.48 [1.41, 1.54] |
| Tersus-buffer1 | 2 | 5504 | 154226 | 1.16 [1.13, 1.19] | 0.87 [0.84, 0.89] | |
| Tersus-buffer2 | 1 | 6282 | 130891 | 1.56 [1.52, 1.60] | 5.1 | 1.31 [1.28, 1.35] |
| Tersus-buffer2 | 2 | 1299 | 30683 | 1.38 [1.30, 1.45] | 1.13 [1.06, 1.19] | |
| TruSeq | 1 | 312 | 14164 | 0.72 [0.64, 0.79] | 4.1 | 0.52 [0.45, 0.58] |
| TruSeq | 2 | 362 | 16705 | 0.70 [0.63, 0.78] | 0.50 [0.44, 0.57] | |
| KTN | 1 | 16802 | 132733 | 4.12 [4.06, 4.17] | 12.92 | 3.49 [3.43, 3.54] |
| KTN | 2 | 6298 | 44331 | 4.62 [4.51, 4.73] | 3.99 [3.89, 4.09] |
*Encyclo (Evrogen JSC), Kapa HiFi PCR Kit (Kapa Biosystems), Phusion High-Fidelity DNA Polymerase (NEB), SD-HS29, SNP-detect, HS Taq, Tersus in buffer 1 (Mg2+ 3.5 mM, Ph 8.5), Tersus in buffer 2 (Mg2+ 2.5 mM, Ph 8.0) (Evrogen JSC), TruSeq Custom Enrichment Kit (Illumina), KTN (KlenTaq N polymerase)30.