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nPfu-Special

No Product Name Cat.# Conc Size Price(₩) Add
1 nPfu-Special P100S 2 unit/ul 200 units ₩ 200,000 추가
2 nPfu-Special P100M 2 unit/ul 400 units ₩ 360,000 추가
3 nPfu-Special P100L 2 unit/ul 1,000 units ₩ 800,000 추가
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Product description
nPfu-Special DNA polymerase was cloned from Pyrococcus furiosus and genetically modified for high processivity. nPfu-Special DNA polymerase is highly efficient in amplifying longer DNA than 10 kb in relatively short period of time (15-30 sec/kb). It has lower error rate of 4.4 X10-7, 50 and 6 times lower than Taq and wild type Pfu DNA polymerase, respectively.

 

Characteristics

- Polymerase speed allows short extension times (15 to 30 s/kb)

- Robust performance, minimal optimization needed

- Can be used for routine PCR as well as long or difficult templates
- Error rate: 4.4 X 10-7
- The blunt end amplicons

 

Applications

- High-performance and high-fidelity PCR

- Cloning (blunt end)

- Difficult (GC-rich) templates

- Template generation for sequencing

- Long-range PCR (>10 kb)

- Mutagenesis

- Microarray

 

Supplied with

- 10X nPfu-Special A buffer

- 10X nPfu-Special B buffer

- 100% DMSO

- dNTP Mixture (2 mM each)

- Sterile water

 

Unit definition 
One unit is defined as the amount of enzyme required to incorporates 10 nmol of dNTP into acid insoluble materials in 30 min at 74
 in 50  reaction mixtures (20 mM Tris-HCl/pH 8.8, 50 mM KCl, 2.5 mM MgCl2, 10 mM 2-mercaptoethanol, 12.5  calf thymus DNA). 

Quality control
- Purity: > 99% on SDS-PAGE
- Endonuclease free
- Exonuclease free

 

Storage buffer
20 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM DTT, 100 mM KCl, 200 
 / ml BSA, 50% glycerol

 

10X nPfu-Special reaction buffer 
- 10X nPfu-Special A buffer

- 10X nPfu-Special B buffer

Buffer A is recommended for a regular PCR, but buffer B can be used when PCR is not successful with 

    buffer A in absence and pressence of DMSO.

  

Standard reaction conditions

- PCR mixturea

10X nPfu-Special buffer

2 μl

nPfu-Special DNA Polymeraseb (2 units/μl)

0.5 μl

dNTP Mixture (2 mM each)

2 μl

Template DNAc (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

bAdd the PCR polymerase at the final step

cPlasmid DNA, 0.1 ng-30 ng; genomic DNA, 50 ng-500 ng

 

- PCR cycle

Step

Temperature

Time

Cycle

Initial denaturation

95

2 min

1

Denaturation

95

10 sec

25~35

Annealing

55~65

5~30 sec

Elongation

72

15~30 sec/kb

Final elongation

72

5 min

1

 Two-step PCR is recommended for longer PCR products than 5 kb. Use 68 for both elongation and annealing temperatures. Reduce the elongation time or amount of enzyme if smeared PCR bands appear. Try reduced amounts of enzyme if the template amount is low. Use DMSO (3~10%) to amplity difficult template targets (GC-rich)

 

● Material Safety Data Sheet

- Enzynomics_MSDS_P100S_nPfu-Special

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