Next-Generation Sequencing (NGS)

High-Specificity Pfu HS Mix

A high-fidelity DNA polymerase with aptamer-based hot-start technology highly suited to target enrichment, NGS library amplification, and cloning applications. High-Specificity Pfu HS Mix provides high yield and sensitivity, as well as superior amplification of GC- and AT-rich targets. The hot-start aptamer binds reversibly to the polymerase which inhibits its activity at ambient temperatures and improves the specificity of the amplification. Product Highlights • Aptamer hot-start for increased specificity • Inhibitor-tolerant (tolerates up to 20% whole blood) PRODUCT CAT NO. VOLUME REACTIONS MDX006 5 mL 200 Rxns

High-Specificity Pfu HS Mix

100 mL 4,000 Rxns

• Robust amplification across different GC-content (29% - 65%) • High-fidelity reduces errors for NGS library amplification • Convenient room temperature reaction set-up • Suitable for multiplexing

Low GC Bias at Both High (over 65%) and Low (under 25%) GC Content

GC bias plot from whole-genome sequencing of a mixture of S. aureus and R. spheroides genomic DNA using High-Specificity Pfu HS Polymerase and supplier K. Libraries were prepared from 100 ng (amplified libraries) or 1 μg (PCR-free library) of genomic DNA from S. aureus and R. spheroides , mixed equimolarly. Library amplification consisted of 10 PCR cycles using High-Specificity Pfu HS Mix (MDX006) or supplier K mix, following the manufacturer’s recommendations. GC bias plots were generated, with %GC content of 100 bp windows on the X axis. Normalized coverage is indicated for the High-Specificity Pfu HS Mix (blue line) and supplier K (green line) and can be compared to a PCR-free reference library (black line), which has the horizontal bias distribution closer to 1 (representing unbiased coverage). The results illustrate High-Specificity Pfu HS Polymerase has a lower bias across the entire range of GC content, compared to the PCR-amplified library from supplier K, especially for GC percentages higher than 65%.

Reference Genome

MDX006

2.0

Supplier K

PCR-Free

1.8

1.6

1.4

1.2

1.0

0.8

0.6

0.4

0.2

0.0

GC% of 100 base windows

Reference Genome

MDX006

Kapa

PCR-Free

Efficient amplification of DNA containing a wide range of GC-content

M 29% 32% 44% 55% 62% 65%

The following regions from human genomic DNA were amplified: RBB8 161 bp (29% GC), RBB8 450 bp (32% GC), IL 10 417 bp (44% GC), Mpex 418 bp (55% GC), EGFR 525 bp (62% GC), EGFR 380 bp (65% GC). Efficient amplification demonstrated across the different GC content tested the suitability of High-Specificity Pfu HS Mix for applications requiring low GC-bias.