PCR simplified with universal annealing
Invitrogen Platinum II Taq Hot-Start DNA Polymerase is designed to get you to your research destination, faster. A universal primer annealing feature reduces optimization steps and allows for co-cycling of all assays. A unique combination of innovative buffer, high-performance Taq DNA polymerase, and superior hot-start technology enables exceptional PCR results, even in the toughest applications.
- Universal primer annealing at 60°C—reduces tedious optimization steps and enables co-cycling of all assays
- 4x faster DNA synthesis and inhibitor resistance—utilizes an engineered Taq polymerase that is extremely robust
- Platinum hot-start technology—offers superior specificity, sensitivity, and yields; allows for room temperature reaction setup
- Green buffer formats—helps reduce pipetting errors with direct gel loading
On this page
- Platinum II Taq Hot-Start DNA Polymerases advantages
- Performance comparison of Platinum II Taq Hot-Start versus Platinum Taq DNA Polymerases
- Ordering information
- Resources–product manuals, education, and support
- Application notes:
Engineered for better PCR
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Platinum II Taq Hot-Start DNA Polymerase advantages
PCR assays using conventional PCR reagents require specific protocols for amplification of each DNA fragment due to varying primer annealing temperatures and different durations of the extension step. Therefore, individual assays cannot be amplified in the same PCR run. With Platinum II Taq Hot-Start DNA Polymerase, different PCR assays can be cycled in parallel using the same protocol with universal primer annealing temperature and the extension step selected for the longest fragment to be amplified. Moreover, Platinum II Taq Hot-Start DNA Polymerase is a “fast” DNA polymerase; thus, the combination of this next-generation DNA polymerase and the universal protocol permits fast cycling of all assays in as little as 30 minutes.
Figure 1. Time saving enabled by assay co-cycling. 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, with traditional PCR reagents, multiple targets often cannot be amplified together in the same PCR run. With Platinum II Taq Hot-Start DNA Polymerase, different PCR assays can be cycled together using one protocol with a universal primer annealing temperature and the extension step selected for the longest fragment to be amplified. Moreover, Platinum II Taq Hot-Start DNA Polymerase is a fast DNA polymerase, delivering PCR results in as little as 30 minutes.
Figure 2. Platinum II Taq Hot-Start DNA Polymerase enables cycling of shorter and longer amplicons together. 132 bp, 251 bp, 1,005 bp, and 3.9 kb fragments were amplified from 50 ng of human genomic DNA in 50 μL reactions using Platinum II Taq Hot-Start DNA Polymerase or other hot-start DNA polymerases: (A) NEB OneTaq Hot Start DNA Polymerase, (B) Qiagen Fast Cycling PCR Kit, (C) Roche FastStart Taq DNA Polymerase. The same protocol was used for all four targets with the annealing and extension settings indicated. The size marker is Thermo Scientific ZipRuler Express DNA Ladder 2.
Platinum II Taq Hot-Start DNA Polymerase is an engineered enzyme with increased DNA synthesis rate. Therefore, with Platinum II Taq Hot-Start DNA Polymerase PCR results are generally more than 2 times faster than other hot-start Taq DNA polymerases.
Figure 3. Fast cycling reduces PCR run time. Amplification of a 529 bp fragment from 50 ng of human genomic DNA in 50 μL reactions for 35 cycles was carried out using Platinum II Taq Hot-Start DNA Polymerase and hot-start DNA polymerases from other suppliers: (A) Sigma-Aldrich KAPA2G Fast HotStart PCR Kit, (B) NEB OneTaq Hot Start DNA Polymerase, (C) Promega GoTaq G2 DNA Polymerase, (D) Toyobo Quick Taq HS DyeMix, (E) Roche FastStart Taq DNA Polymerase, and (F) Sigma-Aldrich JumpStart Taq DNA Polymerase. Cycling times for each polymerase are shown in purple, while ramping times on the ProFlex PCR System (6°C/sec peak block ramp rate) are shown in red. PCR product analysis in 1% TAE agarose gels is presented below the graph. The size marker is the ZipRuler Express DNA Ladder 2.
The high sensitivity of Platinum II Taq Hot-Start DNA Polymerase enables successful amplification of specific product in experiments where there is a limited amount of starting material or the target DNA is in low concentration in the sample.
Figure 4. High sensitivity and reliable amplification from low amounts of input DNA. Amplification of a 529 bp fragment from 0 (no template control); 0.016; 0.08; 0.4; 2; 10; 50; 250 ng of human genomic DNA were amplified in 50 μL PCR reactions using Platinum II Taq Hot-Start DNA Polymerase or competitor DNA polymerases (A—KAPA2G Fast HotStart, B—NEB OneTaq Hot Start, C—Promega GoTaq G2, D—Sigma JumpStart Taq, and E—Takara Taq HS Perfect Mix). The estimated copy number is ~5 copies per 0.016 ng of human genomic DNA. The molecular weight marker is ZipRuler Express DNA Ladder 2.
Platinum II Taq Hot-Start DNA Polymerase was engineered for resistance to inhibitors and helps enable successful amplification with samples of suboptimal purity.
Figure 5. Resistance to inhibitors. Amplification of a 1 kb fragment from human genomic DNA using Platinum II Taq Hot-Start DNA Polymerase or competitor DNA polymerases (A—KAPA 2G Robust HotStart, B—NEB OneTaq Hot Start, C—Promega GoTaq G2, and D—Takara Taq Hot Start Version) in reaction mixtures containing: 1—humic acid (up to final concentration of 1.3 µg/mL), 2—hemin (up to final concentration of 6 µM), 3—xylan (up to final concentration of 0.26 mg/mL), or 4—no inhibitor control. The molecular weight marker is ZipRuler Express DNA Ladder 2.
Figure 6. Amplification of DNA extracted from FFPE tissue samples. Amplification of a 527 bp fragment from varying amounts of DNA extracted from mouse FFPE tissue samples using Platinum II Taq Hot-Start DNA polymerase. RecoverAll Total Nucleic Acid Isolation Kit for FFPE was used for DNA extraction. NTC: no template control. PC: positive control from 1 ng of purified mouse genomic DNA. The molecular weight marker is ZipRuler Express DNA Ladder 2.
The formulation of Platinum II Taq Hot-Start DNA Polymerase and 2X Master Mixes allows for amplification of versatile range of targets, from AT-rich to GC-rich. A separate vial of Platinum GC Enhancer is provided for specific amplification and improved yields of targets with high-GC content.
Figure 7. Robust amplification of AT-rich and GC-rich targets. Various DNA fragments of increasing GC content (indicated above the corresponding lanes) were amplified from 100 ng of human genomic DNA in 50 µL PCR reactions. Platinum GC Enhancer was used for targets with >65% GC. The molecular weight marker is ZipRuler Express DNA Ladder 2.
PCR fragments generated by Platinum II Taq Hot-start DNA Polymerase work well for Sanger sequencing. The enzyme’s superior performance, universal primer annealing, and fast synthesis enable generation of PCR amplicons for Sanger sequencing, with ease and simplicity.
Figure 8. High-quality Sanger sequencing results. A 1.6 kb PCR fragment amplified by Platinum II Taq Hot-start DNA Polymerase was Sanger sequenced using Applied Biosystems 3130xl Genetic Analyzer. Data reported by the KB basecaller of the built-in sequencing analysis software is shown. Clear-range read length (CRL) is defined as the longest uninterrupted segment of bases at a given Quality Value (QV). QV20 corresponds to 1% probability of a base call error and QV30 corresponds to 0.1%. QV>20 is considered high quality and acceptable in most cases.
What is the difference between Platinum II Taq Hot-Start DNA Polymerase and Platinum Taq DNA Polymerase?
Platinum Taq DNA Polymerase has been trusted by researchers for over two decades and has been used in several thousand publications. Platinum II Taq Hot-Start DNA Polymerase is the next generation hot-start DNA polymerase, newly engineered for rapid, robust performance. We recommend that investigators starting new projects use Platinum II Taq DNA Polymerase so that they can benefit from its superior performance, summarized in the table below.
Comparison of technical specifications between Platinum II Taq Hot-Start and Platinum Taq DNA Polymerases
|Platinum II Taq Hot-Start DNA Polymerase||Platinum Taq DNA Polymerase|
|Universal annealing protocol||Yes||No|
|Speed||15 sec/kb||1 min/kb|
|Flexible extension stepa||Yes||No|
|Target length||Up to 5 kb||Up to 5 kb|
|Fidelity versus Taq DNA Polymerase||1x||1x|
|Blunt or 3’A end||3’A||3’A|
|Benchtop stability of assembled PCR reactions||24 h||24 h|
|Controlled low level of residual human and bacterial DNAd||Yes
(≤1 copy of bacterial
aThe extension step can be extended up to 60 sec/kb without the effect on specificity bDirect gel loading with green buffer options. cGreen buffer available as separate item for use with stand-alone enzyme dDuring manufacturing of Platinum II Taq Hot- Start DNA Polymerase, strict measures are taken to control and verify that no more than one copy of residual bacterial genomic DNA is present per unit of the polymerase.
Platinum II Taq Hot-Start DNA Polymerase
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