Bio-Helix NanoTaq II is an advanced hot-start enzyme DNA Polymerase developed with nano technology, building on the reliable performance of the original NanoTaq. It provides enhanced robustness and reliability for your PCR applications. NanoTaq II supports reactions at room temperature and cycling conditions using the same protocol as conventional Taq, while providing reliable amplification from low amounts of input DNA, efficient amplification across a broad range of GC content, and reduced nonspecific amplification. Compared with the original NanoTaq, NanoTaq II offers improved inhibitor resistance, stronger amplification signals, and enhanced amplification of long DNA fragments up to 5 kb.

Figure 1. Comparison of non-specific amplifying effects to major suppliers.
All amplifications were performed in accordance with the manufacturer’s instructions to amplify 320 to 941 bp amplicons from human genomic DNA. nanoTaq II provided both higher specificity and yield products with least band shifting compared to other commercial antibody-mediated hot start polymerases. Lane M: 1Kb plus DNA Ladder RTU (DM015-R500), Lane A: Nano Taq, Lane B: Nano Taq II, Lane C: Thermo DreamTaq, Lane D: Thermo DreamTaq HS, Lane E: Roche KAPA2G HS, Lane F: Invitrogen Platinum HS

*Thermo DreamTaq, Thermo DreamTaq HS and Invitrogen Platinum HS are registered trademarks of Thermo Scientific^TM Pierce^TM. Roche KAPA2G HS is a registered  trademark of Merck. The trademark holders are not affiliated with Bio-Helix Co., Ltd. and do not endorse these products.

Figure 2. Sensitivity and reliable amplification from low amounts of input DNA.
Amplification of a 803 bp fragment from 3, 0.3; 0.2; 0.1; 0.03; 0(no template control) ng of human genomic DNA were amplified in 20μL PCR reactions using nanoTaq DNA Polymerase.

Figure 3. Efficient amplification of DNA sequences with a range of GC content.

A series of DNA fragments of increasing GC content were amplified from human gDNA. nanoTaq Hot-Start DNA Polymerase was used for targets with >50% GC. Lane M: 1Kb plus DNA Ladder RTU (DM015-R500), Lane A: Nano Taq (DP001-0100), Lane B: Nano Taq II (DP003-0100)

Figure 4. Inhibitor resistance on nanoTaq DNA polymerase
A 280 bp human genomic DNA target was conducted to compare with four leading competing DNA polymerases. All amplifications were performed in accordance with manufacturer’s instructions where the mixtures contain:

Lane 1. no inhibitor control
Lane 2. bile salts with final concentration of 0.5 mg/mL
Lane 3. heparin with final concentration of 0.5 unit/mL
Lane 4. sodium citrate with final concentration of 150 μg/mL
Lane 5. hemin with final concentration of 2 μg/mL
Lane 6. humic acid with final concentration of 8 μg/mL

The results show that nanoTaq has the best tolerance with the humic acid interference, when compared with major suppliers of the hot-start DNA polymerase.

*Thermo DreamTaq, Thermo DreamTaq HS and Invitrogen Platinum HS are registered trademarks of Thermo Scientific^TM Pierce^TM. Roche KAPA2G HS is a registered trademark of Merck. The trademark holders are not affiliated with Bio-Helix Co., Ltd. and do not endorse these products.

Figure 5. Amplification of long target DNA fragments
It is generally known that Taq DNA polymerase exhibits difficulty amplifying DNA fragment sizes larger than 1 kb. This is probably due to the polymerase’s error-prone amplification process which reduces the efficiency in amplifying longer target sizes. However, we have shown that nanoTaq is capable of amplifying. Lane M: 1Kb plus DNA Ladder RTU (DM015-R500), Lane 1: 0.5 kb, Lane 2: 1 kb, Lane 3: 2 kb, Lane 4: 3 kb, Lane 5: 5 kb and Lane 6: 8 kb Lambda DNA fragment sizes under the 5-min extension time at 72°C. Therefore, nanoTaq II shows optimal performance in applications with long PCR fragments.

*Invitrogen Platinum HS are registered trademarks of Thermo Scientific^TM Pierce^TM. The trademark holders are not affiliated with Bio-Helix Co., Ltd. and do not endorse the product.

► High tolerance aganist various reaction inhibitors
► Reduce nonspecific primer annealing
► Plays in efficient PCR amplification of GC-rich sequences
► Using the same protocol and cycling conditions as conventional Taq DNA polymerases

1. For each 20 μl reaction, assemble the following in a 0.2 ml PCR tube on ice just before use:

2. Mix gently. If necessary, centrifuge briefly and cap tubes.

3. Place them into the thermocycler and process for 30-35 cycles as follows:

Note: Optimal conditions for amplification will vary depending on the primers and thermal cycler used. It may be necessary to optimize the system for individual primers, template, and thermal cycler.

* Template:
Purified high quality DNA is needed for a success PCR reaction. The final concentration of cDNA template, please refer to “Reaction Setup”.

* Storage Buffer:
The enzyme is supplied in a storage buffer consisting of 20 mM Tris-HCl (pH 7.5), 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 50% (v/v) glycerol, and 0.5% Tween 20.

* Unit Definition:
One unit is defined as the amount of enzyme that will catalyze the incorporation of 10 nmol of dNTP into acid- insoluble form in 30 min at 74°C in a reaction containing 25 mM TAPS (tris-[hydroxymethyl]-methyl-amino- propane-sulfonic acid, sodium salt), pH 9.3 at 25°C, 50 mM KCl, 2 mM MgCl₂, 1 mM β-mercaptoethanol, 0.2 mM dATP, dGTP, and dTTP, 0.1 μM [α-32P] dCTP, and activated salmon sperm DNA.