Hot-start formulations use an antibody, aptamer, or chemical modification to block polymerase activity at room temperature. Activity is restored only during the initial high-temperature denaturation step. This prevents non-specific extension of mis-annealed primers during reaction setup. It reduces primer-dimer formation and non-specific bands, which is especially helpful with degenerate primers or complex genomic templates.
PCR & qPCR
High Fidelity PCR Polymerase and PCR Master Mixes for Every Amplification
High fidelity PCR polymerase and PCR master mixes for every amplification
Get the right PCR enzyme or master mix for your job, from high fidelity polymerases for cloning and sequencing to ready-to-use mixes for fast routine amplification. Your options span a broad range of formats and fidelity levels. Your DNA polymerase choice determines error rate and processivity, and whether your protocol can tolerate inhibitors or complex templates.
Standard Taq-based master mixes run most of your routine PCR. Ready-to-use 2x formulations offer high sensitivity and a broad amplification range. Hot-start variants are available to reduce non-specific amplification when cycling from cold. Hot-start formats use antibody or aptamer blockers that stop polymerase activity until the first denaturation step. Loading-dye-incorporated mixes let you load the gel directly, without a separate buffer addition.
For your high fidelity applications, proofreading polymerases carry 3-prime to 5-prime exonuclease activity. This gives error rates well below standard Taq. Hot-start high fidelity options are available for your challenging templates.
Direct PCR formats amplify from blood and crude lysates without a separate extraction step. They are formulated to tolerate haemoglobin and many common inhibitors under recommended conditions. Phi29 DNA Polymerase supports isothermal whole-genome amplification and rolling-circle amplification from your low-input samples at constant temperature.
Match the polymerase to your application
1) Routine colony PCR or research screening. A standard Taq 2x master mix suits this work. It is cost-effective for your high-volume screening, where cloning accuracy is not critical.
2) Hot-start for specificity. A hot-start 2x master mix helps when non-specific bands appear in your gel. Antibody or aptamer blockers prevent polymerase activity until the first denaturation step.
3) High fidelity for cloning or sequencing. A proofreading polymerase mix with 3-prime to 5-prime exonuclease activity suits this application. Hot-start high fidelity options are available for long or GC-rich amplicons over 5 kb.
4) Amplification from crude or inhibitor-rich samples. A direct PCR master mix formulated for blood lysates or crude tissue extracts suits this work. These formulations are designed to tolerate haemoglobin and many common PCR inhibitors without a prior extraction step, when used as directed.
5) Whole-genome or rolling-circle amplification. Phi29 DNA Polymerase at 30 degrees C suits this need. It displaces strands without thermal cycling and generates high-molecular-weight product from your nanogram input.
Applications
Routine colony and research screening
Standard Taq mixes support your routine colony screening and research genotyping, a cost-effective choice for high-volume PCR. Pair with PCR accessories.
Hot-start specificity and multiplex genotyping
Hot-start formulations improve specificity in your complex templates and multiplex genotyping panels, cutting primer-dimer and non-specific bands.
Cloning and sequencing template preparation
Proofreading polymerases suit your cloning or sequencing amplicons, cutting error rates 10 to 100-fold versus Taq. See molecular cloning kits.
Long-range and GC-rich amplification
Long-range formulations blend Taq with a proofreading enzyme to amplify your templates up to 20 kb, beyond standard high-fidelity mix reach.
Crude-sample amplification
Direct PCR mixes amplify crude blood, swabs, or tissue punches without extraction, speeding screening in research, plant, and animal genotyping laboratories.
Whole-genome and rolling-circle amplification
Phi29-based amplification handles your limited material for single-cell and archival samples. See sample preparation kits.
Frequently asked questions
Taq polymerase lacks a 3-prime to 5-prime exonuclease domain. It has an error rate of roughly one substitution per 10,000 to 100,000 bases. Proofreading polymerases carry a 3-prime to 5-prime exonuclease that excises misincorporated bases, reducing the error rate ten-fold or more. Proofreading enzymes also produce blunt-ended products rather than the single-base 3-prime adenine overhang characteristic of Taq. This affects downstream cloning strategy.
Specialised direct PCR master mixes are formulated with additives that neutralise common inhibitors in blood, including haemoglobin, immunoglobulins, and heparin. You add a small volume of lysed blood or crude tissue extract directly to the reaction. This approach is well-established for genotyping applications. It may not provide sufficient sensitivity for low-copy targets, where full extraction and purification are still preferred.
Standard Taq-based mixes reliably amplify up to 2 to 3 kb. Proofreading mixes extend this to 5 to 10 kb for most templates. Long-range formulations blending Taq and a proofreading enzyme can amplify 20 kb or more. GC content, secondary structure, and template quality all limit achievable amplicon length in practice.
Product catalogs
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