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RbTaq™ Tenuto HOT new

No Product Name Cat.# Conc Size Price(₩) Add
1 RbTaq™ Tenuto HOT P225RB 5 units/μl 250 units ₩ 104,000 추가
2 RbTaq™ Tenuto HOT P250RB 5 units/μl 500 units ₩ 187,000 추가
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Product description

RbTaq™ Tenuto HOT is a repebody mediated hot-start Taq DNA polymerase with 3'→5’ proofreading activity. RbTaq™ Tenuto HOT can be used to amplify DNA longer than 10 kb, which is difficult with common Taq polymerases. This product is improved in both fidelity (> 2 fold) of PCR products and amplification efficiency of longer PCR products. RbTaq™ Tenuto HOT is combined with a repebody that inactivates the enzyme until the first denaturation step. Thus, RbTaq™ Tenuto HOT is suitable for high-speed PCR and designed to have enhanced specificity, sensitivity and amplification efficiency of various targets.

 

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

 

Supplied with

- 10X RbTaq™ Tenuto HOT Buffer

- 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).

 

Storage buffer

20 mM Tris-HCl (pH 7.9), 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.5% NP-40, 0.5% Tween-20, 50% glycerol.

 

GC Melt I and II

- This product is useful for amplification of DNA with GC-rich 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.

 

Standard reaction conditions

- PCR mixturea

10X RbTaq™ Tenuto HOT Buffer

2 μl

RbTaq Tenuto HOT (5 units/μl)

0.2 μl

dNTP Mixture (2 mM each)

2 μl

Template DNAb (0.1~500 ng/μl)

1 μl

Primer 1 (5 pmole/μl)

1 μl

Primer 2 (5 pmole/μl)

1 μl

Sterile water

up to 20 μl

aAssemble the reaction mixture on ice

bPlasmid DNA, 0.1 ng~30 ng; genomic DNA, 50 ng~500 ng

 

- PCR cycle

Step

Temperature

Time

Cycle

Initial denaturationa

95

3 min

1

Denaturation

95

30 sec

25~35

Annealingb

55~65

30~60 sec

Elongation

72

1 min/kb

Final elongation

72

5 min

1

When cycles are over, keep the reaction mixture at 4; may add 10 mM EDTA until use to prevent DNA degradation.

a30 sec at 95 for cDNA and 3 min at 95 for genomic DNA is recommended for enzyme activation.

bRecommended annealing temperatures is 5 to 10 below the lower Tm of the two primers used.




Figure 1. Seven different genes were amplified using 10 ng of HeLa gDNA as a template to compare amplification efficiency and specificity. Seven primer sets difficult to amplify were selected

● Material Safety Data Sheet

- Enzynomics_MSDS_P225RB_RbTaq™ Tenuto HOT

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