Close

Glycerol-Free High-Fidelity Pfu (HC)

PCR is used for the amplification of a next-generation sequencing (NGS) library. In addition to increasing the amount of DNA, the PCR amplification step is also used for DNA target enrichment for fragments that have an adaptor-ligated to each end. These will be the fragments that attach to the flow cell and will be sequenced, so it is important that this PCR amplification step does not introduce mutations to the NGS library, as a lower error rate means that less sequence coverage is required, saving expense and time.

Request a Sample


Have questions about a product?

Contact us to learn more about Meridian’s molecular or immunoassay reagent portfolio. We want to hear from you!

Contact A Specialist

Meridian's Glycerol-Free High-Fidelity Pfu (HC)

  • High concentration Pfu polymerase allowing for bulk production of PCR master mixes

  • An error rate of 3.0 x 10-6

  • Ideal for NGS library amplification

  • Glycerol-free for preparation of dried-down PCR tests

Multiplex amplification

Glycerol-Free High-Fidelity Pfu (HC) graph
Glycerol-Free High-Fidelity Pfu (HC), was lyophilized with Lyo-Ready Pfu Reaction Buffer, 5x and stored at 37°C for 7 and 28 days. Lyophilization samples were tested in multiplexed PCR amplification of 7 targets (793 bp, 649 bp, 548 bp, 418 bp, 332 bp, 196 bp, 135 bp) against the same enzyme and reaction buffer without lyophilization (Wet Reference) to demonstrate stable after lyophilization allowing for extended storage periods and use in NGS library amplification.

Glycerol-Free High-Fidelity Pfu (HC), MDX203

Application: PCR Specimen Type: DNA Polymerase,NGS Enzymes Concentration: 20 U/µL High Concentration: Yes Wet: Yes
Glycerol-Free: YesYes Lyo-Ready: Requires excipients Air-Dryable: Requires excipients Dryable: Requires excipients Sustainability: Glycerol-Free

Glycerol-free, high concentration, high fidelity DNA polymerase with separate dilution and reaction buffer, ideal for NGS library amplification and gene expression. Ideal for lyophilization.

Available in 0.025 mL (500 Units) or 1 mL (20,000 Units) aliquots

Description

High-Fidelity Pfu is a thermostable high fidelity DNA polymerase enzyme, isolated from Pyrococcus furiosus. The Pfu polymerase enzyme replicates DNA in the 5´→3´ direction in the presence of magnesium, but also possesses 3´→5´ exonuclease (proofreading) activity. Base misincorporations that may occur during polymerization are rapidly excised by this proofreading activity. High-Fidelity has an error rate of 3.0 x 10-6 generating blunt-ended amplicons up to 5 kb in length. Glycerol-Free High-Fidelity Pfu (HC) is supplied in a glycerol-free storage buffer and is accompanied by a 5x Reaction Buffer that contains magnesium, dNTPs and excipients required for lyophilization.


Specifications

Description High concentration, high-fidelity Taq DNA polymerase with separate Pfu Polymerase Dilution Buffer and10x Pfu Reaction Buffer.
Concentration 20 U/µL
Appearance Clear, colorless solution
Application PCR, NGS library preparation, cloning, protein expression
Sample type cDNA, DNA
Presentation 3 vials
Storage -20 °C
Mix stability See outer label
Consistency ± 0.5 Ct variance between test and reference sample
DNA Contamination None detected in PCR amplification with traces overlay with the negative control on E. coli and mouse genomic DNA specific targets.
DNase Contamination No detectable degradation

FAQs: High-Fidelity Pfu

In nature, the ability of a DNA polymerase to correct misincorporation of nucleotides in the DNA strand being elongated is often crucial to the survival of the host organism. This ability is termed “proofreading activity” and occurs in the 3′ to 5′ direction. This activity also leads to the polymerase removing unpaired nucleotides overhanging at the 3’end (A-overhangs), creating blunt ends.

If possible, we would recommend an additional clean-up of affected samples, alternatively High-Specificity Pfu HS Mix (MDX006) would be recommended, as it is designed to work in the presence of inhibitors.

High-Fidelity Pfu has been validated for templates up to 5 kb.

DNA target enrichment is a pre-sequencing DNA preparation step where DNA sequences are either directly amplified (multiplex PCR-based) or captured (hybrid capture-based). These enriched DNA fragments can then be sequenced using DNA sequencers.

A lyophilized PCR molecular test will remain stable for up to 2 years at ambient temperature if correctly stored in sealed pouches.

Get In Touch With A Specialist

Have questions about a product? Want to learn more about Meridian’s molecular or immunoassay reagent portfolio? We want to hear from you!


By submitting your information in this form, you agree that your personal information may be stored and processed in any country where we have facilities or service providers, and by using our “Contact Us” page you agree to the transfer of information to countries outside of your country of residence, including to the United States, which may provide for different data protection rules than in your country. The information you submit will be governed by our Privacy Statement.

Learn More About Us

Learn More
Welcome to
logo blue

Please select your country to continue

Continue
CN