SKU : P225A
หมวดหมู่ : PCR Enzyme / RT-PCR ,  2. Cell & Molecular Biology ,  Real-Time PCR ,  Enzynomics (Korea) , 
แบรนด์ : Enzynomics (South Korea)
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Product description
nTaq-Tenuto is nTaq (Cat. P025, P050) supplemented with 3'->5’ proofreading activity and a PCR enhancing factor for improved efficiency and fidelity. nTaq-Tenuto can be used to amplify DNA longer than 10 kb, which is difficult with common Taq polymerases alone. Thus, this product is improved in both fidelity (> 2 fold) of PCR products and amplification efficiency of longer PCR products.
Characteristics
- Molecular weight: 94 kDa
- Error rate: 3.0 X 10-6
- Thermal stability: Half life of 40 min at 95℃
- A-tail formation at 3’ ends of amplified DNA products.
Applications
- Amplification of long DNA fragments (<10 kb)
- Amplification of high-complexity template DNA
- Primer extension
- Colony PCR
- Labeling of DNA fragments with radioactive-isotopes
- Nucleotide sequencing
Supplied with
○ nTaq-Tenuto (Mg2+ plus buffer)
- 10X nTaq-Tenuto buffer (Mg2+ plus)
- dNTP Mixture (2 mM each)
- GC Melt I
- GC Melt II
- Sterile water
Quality control
- Purity: >99% on SDS-PAGE
- Endonuclease-free
- Exonuclease-free
- RNase free
- Inhibitor-free
Unit definition
One unit is defined as the amount of enzyme required to incorporate 10 nmol of dNTP into acid insoluble materials in 30 min at 74℃ in a 50-μl reaction mixture (20 mM Tris-HCl/pH 8.8, 50 mM KCl, 2.5 mM MgCl2, 10 mM β-mercaptoethanol, 12.5 μg of calf thymus DNA).
10X nTaq-Tenuto buffer
Mg2+ plus buffer: Containing 20 mM MgCl2
GC Melt l and ll
- This product is useful for amplification of DNA with GCrich sequences or to avoid amplification of non-specific bands (Use of GC Melt may reduce PCR efficiency).
- In general, use 1X strength in PCR reaction by diluting the 10X solution, but the amount should be adjusted for optimal results.
Figure 1. PCR amplification of long DNA fragments using nTaq-Tenuto.
DNA fragments up to 20 kb can be efficiently amplified by nTaq-Tenuto (Cat.# P225 or P250). Target DNAs with varying lengths were amplified by the use of following PCR cycles; 95℃ 2 min, (95℃ 30 sec, 55℃ 30 sec, 72℃ 1 min/kb) x 30, 72℃ 5 min. Note that elongation time increases by 1 min per kb. Lane 1, 0.5 kb; lane 2, 1 kb; lane 3, 2 kb; lane 4, 3 kb; lane 5, 4.5 kb; lane 6, 8 kb; lane 7, 11 kb; lane 8, 20 kb; lane 9, 1 kb (+) Ladder Marker (Cat.# DM003)