Platinum II Taq Hot-Start DNA Polymerase

Engineered enzyme for high specificity and yield with fast cycling

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 different 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.

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Highlights

  • Universal primer annealing at 60°C—helps simplify optimization steps and enables co-cycling of multiple assays
  • 4x faster DNA synthesis and high inhibitor tolerance—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—help reduce pipetting errors with direct gel loading

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.

Platinum2TaqHot-StartDNA-Polymerase-cycle-together

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.

Time saving enabled by assay co-cycling

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.

Fast cycling reduces PCR run time

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. 

High sensitivity and reliable amplification from low amounts of input DNA

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.

Resistance to inhibitors

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.

Amplification of DNA extracted from FFPE tissue samples

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.

Robust amplification of AT-rich and GC-rich targets

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.

High-quality Sanger sequencing results

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 hot-start PCR?

Learn about common issues in PCR amplification and how you can resolve them with hot-start PCR. Also discover different types of hot-start enzyme modifications and how to choose a hot-start DNA polymerase suitable for your PCR.

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
Inhibitor tolerance Yes No
Target length Up to 5 kb Up to 5 kb
Hot-start modification Antibody-mediated Antibody-mediated
Fidelity versus Taq DNA Polymerase 1x 1x
Amplicon overhangs 3’A 3’A
Benchtop stability of assembled PCR reactions 24 h 24 h
GC-rich amplification Yes Yes
Certified low level of residual human and bacterial DNAd Yes
(≤1 copy of bacterial gDNA/enzyme unit)
No
Master mix formats Colorless/greenb Colorless/greenb
Stand-alone enzyme formats Colorlessc Colorless/greenb

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 for direct loading gel. dDuring manufacturing of Platinum II Taq Hot-Start DNA Polymerase, strict measures are taken to control and verify by qPCR that no more than one copy of residual bacterial genomic DNA is present per unit of the polymerase.

Ordering information

Platinum II Taq Hot-Start DNA Polymerase

Selected references

Applications Product format used Publications
Study of genes in cultures of human and mouse pluripotent stem cells—cDNA was amplified by two-step PCR for a number of genes, then visualized with the SYBR Safe gel stain. Platinum II Taq Hot-Start DNA Polymerase Esseltine JL, Brooks CR, Edwards NA et al. (2020) Dynamic regulation of connexins in stem cell pluripotency. Stem Cells 38:52–66.
Cloning of the sea lamprey GHR gene—GHR was amplified by RT-PCR and TA-cloned. Platinum II Taq Hot-Start DNA Polymerase, supplemented with the included GC Enhancer Gong N, Ferreira-Martins D, McCormick SD et al. (2020) Divergent genes encoding the putative receptors for growth hormone and prolactin in sea lamprey display distinct patterns of expression. Sci Rep 10(1):1674.
Amplification of GC-rich (~70%) chromosomal DNA or cDNA— Multiple amplicons were co-cycled following a universal protocol. Platinum II Hot-Start PCR Master Mix, supplemented with the included GC Enhancer Hackl T, Martin R, Barenhoff K et al. (2019) Four high-quality draft genome assemblies of the marine heterotrophic nanoflagellate Cafeteria roenbergensis. bioRxiv 751586.
Methylation analysis of genomic DNA from colon cells—Bisulfite-treated DNA was amplified and Sanger sequenced. Platinum II Taq Hot-Start DNA Polymerase Ibrahim ML, Klement JD, Lu C et al. (2018) Myeloid-Derived Suppressor Cells Produce IL-10 to Elicit DNMT3b-Dependent IRF8 Silencing to Promote Colitis-Associated Colon Tumorigenesis. Cell Rep 25(11):3036–3046.
Detection of microRNA‑21 expression levels—qPCR SYBR Green assays were performed using cDNA templates. Platinum II Taq Hot‑Start DNA Polymerase Wei X, You X, Zhang J et al. (2019) miR‑21 inhibitor facilitates the anticancer activity of doxorubicin loaded nanometer in melanoma. Oncol Rep 42(1):414–424.
Amplification of the ITS1 region of Sarcocystis genomic DNA—Sanger sequencing was performed afterward. Platinum II Hot-Start Green PCR Master Mix Prakas P, Kirillova V, Gavarāne I et al. (2019) Morphological and molecular description of Sarcocystis ratti n. sp. from the black rat (Rattus rattus) in Latvia. Parasitol Res 118(9):2689–2694.
RT-PCR verification of Sendai reprogramming vector clearance in a transfected iPSC line—Analysis was done on E-Gel agarose gels. Platinum II Hot-Start PCR Master Mix Hong J, Xu M, Li R et al. (2019) Generation of an induced pluripotent stem cell line (TRNDi008-A) from a Hunter syndrome patient carrying a hemizygous 208insC mutation in the IDS gene. Stem Cell Res 37:101451.

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