I-5™ Hi-Fi DNA Polymerase

Description

General Description:
I-5 Hi-Fi DNA polymerase is an ultra-fast and high-fidelity DNA polymerase. It provides robust amplification of from different templates including plasmids, BACS, genomic DNA, and lambda DNA. It allows for amplification of up to 8kb with human genomic DNA and up to 21kb with lambda DNA. It has an extension speed of 1 kb / 10-15 seconds depending on template type. This allows the user to save time by speeding up PCR reactions and provides higher fidelity than Taq or Pfu.

Features:
• Robust – maximal success and minimal optimization needed
• Fidelity – 50X greater than Taq
• High Speed –10X faster than Pfu
• High Yield
• Versatile – best for long or difficult templates

Source:
expressed in E. coli

Applications:
PCR
Cloning
Long or Difficult Amplification
High-Throughput PCR

Supplied with:
5X Reaction Buffer (1ml), 50mM MgCl₂ (1ml)

Supplied in (buffer description):
20mM Tris-HCl, 100mM KCl, 1mM DTT, 0.1mM EDTA, 200ug/ml BSA, 50% glycerol, 1X stabilizer, pH 7.4 @ 25 C

Storage Condition:
-20°C

Unit Definition:
One unit is the amount of enzyme that incorporates 5 nmol of dNTPs into acid insoluble material in 15 minutes at 72°C.

References:
Frey, B. and Suppman, B. (1995). BioChemica. 2, 34-35.
Chester, N. and Marshak, D.R. (1993). Analytical Biochemistry. 209, 284-290.

Shipping Condition:
Dry ice

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).

Conclusion:

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).

Additional information

The Q&A for this product will be available soon.