
I-5 Hi-Fi DNA HotStart Polymerase is a high-fidelity and fast DNA polymerase.
General Description
Source Applications
Supplied with 50mM MgCl2 (1ml) (for optimization)
Supplied in (buffer description)
Storage Condition
Unit Definition Experimental Data High Fidelity DNA Polymerase Comparison of I5 (MCLAB), KAPA (Roche), and Q5 (NEB) DNA polymerase fidelity is estimated by the error rate, i.e., the percentage of base substitutions per PCR cycle (given a replication efficiency of 2.0). The error rates of DNA Polymerase I5 (MCLAB), KAPA (Roche) and Q5 (NEB) are measured by both Sanger sequencing and Next Generation Sequencing (Illumina_MiSeq) (Table 1).
The fidelity of MCLAB’s I5 DNA polymerase is ultra-high, with an average of 1.25 errors per 100,000bp from both Sanger Sequencing and NGS_Illumina, which is at least equivalent to that of KAPA or Q5. Methods: In this assay, I5 was examined to determine its fidelity compared to KAPA and Q5 DNA polymerase. A 2000 bp template was PCR amplified with either I5, or KAPA, or Q5 DNA Polymerase for 35 cycles. The PCR error rates were determined by Sanger sequencing (Figure 1) and NGS_Illumina (Figure 2). The raw data was normalized by the following equation: e = (f⋅2) / n where f is the percentage of errors measured after PCR, and n is the number of actual PCR cycles.
Figure 1. PCR polymerase fidelity estimation by Sanger Sequencing.
Figure 2. PCR polymerase fidelity estimation by Next Generation Sequencing (Illumina_MiSeq).
Reference
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