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TOPsimple™ DryMIX–Forte

No Product Name Cat.# Conc Size Price(₩) Add
1 TOPsimple™ DryMIX–Forte (20 μl reactions) P581F - 96 tubes ₩ 100,000 추가
2 TOPsimple™ DryMIX–Forte (20 μl reactions) P582F - 960 tubes ₩ 800,000 추가
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Product description

This product is formulated for the maximal stability of nPfu-Forte that has been dried together with reaction buffer, dNTP mixture and a stabilizer. This product is ready to use upon the addition of primers and template DNA in distilled water to each tube.

 

Characteristics

- High fidelity PCR due to proofreading activity. This product is suitable for site-directed mutagenesis

or cloning that requires faithful DNA synthesis.

- Simplicity: The dried pellet in TOPsimple™ DryMIX is readily soluble upon addition of primers and

template DNA in water. There is no need of a step to solubilize by mixing.

- Convenience: Dry pellets of TOPsimple™ DryMIX are tightly bound to the bottom of the tubes; easy

to transport and handle.

- Stability: The presence of a unique stabilizer ensures great stability of PCR DNA polymerase even

at room temperature (up to 2 months).

- 2-Dye system: Easy for gel electrophoresis (xylene cyanol and Orange G)

- Blunt end formation at 3’ ends of amplified duplex DNA

 

Applications

- High fidelity DNA amplification for cloning

- Amplification of long DNA fragments (<10 kb)

- DNA amplification to produce blunt end products

- Site-directed mutagenesis

- Colony PCR

 

Components

nPfu-Forte DNA polymerase: 2 units/tube

nPfu-Forte buffer (containing 2 mM MgSO4)

- dNTP mixture: 0.2 mM each

- Stabilizer

- Dyes: Xylene cyanol and Orange G

 

Migration of dyes in agarose gel

In an ordinary agarose gel, xylene cyanol co-migrate with 4-kb DNA fragments, and Orange G with 50-bp DNA fragments.

 

Storage

Stable up to 2 years at -20, 6 months at 4, or 2 months at room temperature (Storage at -20 is recommended).

 

Standard reaction conditions

- PCR mixture

TOPsimple™ DryMIX-Forte

1 tube

Template DNAa (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

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

 

- PCR cycles

Step

Temperature

Time

Cycle

Initial denaturation

95

2 min

1

Denaturation

95

30 sec

25~35

Annealinga

55~65

30~60 sec

Elongation

72

1 min/kb

Final elongation

72

5 min

1

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

aRecommend annealing temperature is 5 to 10 below the lower Tm of the two primers used.

 


● Material Safety Data Sheet

- Enzynomics_MSDS_P581F_TOPsimple™ DryMIX–Forte

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