Invitrogen™ PCR Platinum™ SuperFi II DNA Polymerase

• Exceptional accuracy—Higher than 300x Taq fidelity as determined by next-generation sequencing
• Simplified workflow—Buffer formulated for primer annealing at 60°C (no need for a T_m calculator)
• Increased PCR success—Robust amplification of GC-rich targets, DNA of suboptimal purity, and long sequences
• Enabled automation—High specificity and benchtop stability for 24 hours after reaction setup, enabled by Invitrogen Platinum hot-start technology
• Reduced pipetting—Master mix options available with or without direct gel-loading dyes

PCR Platinum™ SuperFi

>300x Taq Fidelity

Platinum SuperFi II DNA Polymerase offers the highest level of confidence for preserving DNA sequence accuracy with its extremely low error rate. Using next-generation sequencing, the relative fidelity of Platinum SuperFi II DNA polymerase was calculated to be >300x that of Taq DNA polymerase.
 

Fidelity comparison across commercially available enzymes relative to Taq enzyme. A 3.9 kb sequence was amplified by PCR using different DNA polymerases, and the resulting PCR amplicons were then fragmented with a MuA transposase. Unique molecular identifiers (UMI), which consist of 12 random nucleotides, were introduced during fragmentation to individually tag each product. After next-generation sequencing, reads were aligned to the correct sequence, grouped by UMI families, and errors were called. Errors were identified only if they were present in all reads in the UMI family; otherwise they were discarded as sequencing errors. The polymerase fidelities were normalized to the Taq DNA polymerase.

High Yield & Specifity with Platinum SuperFi II PCR DNA Polymerase Kit

Platinum SuperFi II DNA Polymerase amplifies a broad range of target lengths with high specificity and yield due to robustness of the enzyme and superior hot-start technology. The antibody-based Platinum hot-start technology inhibits enzyme activity until the initial PCR denaturation step, preventing nonspecific amplification and primer degradation while allowing greater yield of the target amplicons.
 

Geniş amplikon uzunlukları aralığında çok yönlülük. Platinum SuperFi II DNA Polimeraz (en soldaki panel), 100 ng insan genomik DNA'sından amplifiye edilmiş 0,3 kb ila 14 kb arasında değişen DNA fragmanları aralığında yüksek özgüllük ve verim sağlar. Aynı hedefler rakip DNA polimerazları kullanılarak da amplifiye edildi: A—Merck KOD Hot Start, B—KAPA HiFi HotStart PCR Kit, C—PrimeSTAR GXL. Moleküler ağırlık markörü TrackIt 1 Kb Plus DNA Ladder'dır.

High Sensitivity

The high sensitivity of Platinum SuperFi II DNA Polymerase enables detection of low-abundance DNA templates with accurate results. High sensitivity is advantageous in experiments where there is a limited amount of starting material or the target DNA is in low concentration in the sample.
 

High sensitivity and reliable amplification from low amounts of input DNA. Platinum SuperFi II DNA Polymerase (far left panel) shows reliable amplification of a 2 kb fragment from 0.4 ng, 2 ng, 10 ng, 50 ng, and 250 ng of human genomic DNA (template amounts are indicated above each lane.). The same target was amplified with competitor DNA polymerases: A—PfuUltra II Fusion Hot Start, B—HotStar HiFidelity DNA Polymerase Kit, C—Expand HiFiPLUS Enzyme Blend. The estimated copy number is ~100 copies per 0.4 ng of human genomic DNA. The molecular weight marker is TrackIt 1 Kb Plus DNA Ladder.

Direct Gel Loading for PCR 

Platinum SuperFi II Green PCR Master Mix offers the convenience of direct gel loading of PCR products, eliminating tedious steps of dye addition to PCR samples and helping reduce pipetting errors. The green buffer is compatible with downstream applications including DNA sequencing, ligation, and restriction digestion.

Invitrogen™ Platinum™ SuperFi II DNA Polymerase

Resistance to Inhibitors

Platinum SuperFi II DNA Polymerase is engineered with a DNA-binding domain resulting in high processivity and increased tolerance to common PCR inhibitors such as hemin (red blood cell component), xylan (plant biopolymer), and humic acid (found in soil).

Platinum SuperFi II DNA Polymerase shows high tolerance to common PCR inhibitors. A 2 kb human genomic DNA fragment was amplified from 50 ng of human genomic DNA using Platinum SuperFi II DNA Polymerase or competitor high-fidelity DNA polymerases: A—Q5 Hot Start High-Fidelity, B—PrimeSTAR GXL, C—Merck KOD Hot Start, and D—KAPA HiFi HotStart PCR Kit in reaction mixtures containing 1—no inhibitor, 2—humic acid (4 µg/mL), 3—hemin (20 µM), or 4—bile salt (1 mg/mL). The molecular weight marker is TrackIt 1 Kb Plus DNA Ladder.

Benchtop Stability

Extended stability of the Platinum SuperFi II DNA Polymerase enzyme at room temperature enables high-throughput applications. Its superior Platinum hot-start technology allows benchtop stability and high specificity of the enzyme.
 

Assembled reactions with Platinum SuperFi II DNA Polymerase are stable room temperature. A 0.5 kb fragment was amplified form 50 ng of human genomic DNA. PCR reactions were set up and left at room temperature for 0 hr and 24 hr before loading to the Applied Biosystems ProFlex thermal cycler. Even after 24 hr of room-temperature setup, Platinum SuperFi II DNA Polymerase (lane P) produces results with high specificity and yields. The same experiment was also performed with competitor DNA polymerases: A—Q5 Hot Start High-Fidelity, B—KAPA HiFi HotStart PCR Kit, C—PrimeSTAR GXL. The molecular weight marker is TrackIt 1 Kb Plus DNA Ladder.

 

Universal Protocol

Due to its innovative reaction buffer, Platinum SuperFi II DNA Polymerase allows for a universal annealing temperature and flexible extension time for co-cycling of all assays.

Time saving and assay co-cycling enabled by universal PCR protocol. PCR assays using conventional PCR reagents require specific protocols for amplification of each DNA fragment because of the different primer annealing temperatures and extension steps; therefore, multiple targets often cannot be amplified together in the same PCR run. With Platinum SuperFi II DNA Polymerase, different PCR assays can be cycled together using one protocol with a universal primer annealing temperature and the extension time selected for the longest fragment to be amplified

Platinum SuperFi II DNA Polymerase enables cycling of shorter and longer amplicons together.  0.7 kb, 2.0 kb, 4.8 kb, and 14 kb fragments were amplified from 100 ng of human genomic DNA using the same protocol for all four targets: 98°C denaturation for 10 sec, 60°C annealing for 10 sec, 72°C extension for 7 min. The extension time was based on length of the longest target.

60°C Primer Annealing with Platinum SuperFi II PCR DNA Polymerase

The unique formulation of the Platinum SuperFi II buffer helps reduce tedious optimization step in PCR. Calculation of primer melting temperatures for the annealing step is no longer required with Platinum SuperFi II DNA polymerase. The innovative buffer formulation enables annealing of primers at 60°C regardless of their sequences that follow general primer design rules. The buffer also allows successful amplification when calculated T_ms are used in the annealing step.

 

Platinum SuperFi II DNA Polymerase produces PCR products with high specificity and yield following the universal annealing temperature at 60°C. Primer sets of varying annealing temperature were used to amplify 12 targets from 50 ng of human genomic DNA at 60°C annealing temperature. The molecular weight marker is Invitrogen TrackIt 1 Kb Plus DNA Ladder. The annealing temperatures stated were calculated using the Tm calculator for Platinum SuperFi DNA Polymerase.

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