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Polymerases

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2X TOPsimple™ DyeMIX (aliquot)–Forte

No Product Name Cat.# Conc Size Price(₩) Add
1 2X TOPsimple™ DyeMIX (aliquot)–Forte
(20 μl reactions)
P561F 2X 96 tubes ₩ 100,000 추가
2 2X TOPsimple™ DyeMIX (aliquot)–Forte
(20 μl reactions)
P562F 2X 960 tubes ₩ 800,000 추가
3 2X TOPsimple™ DyeMIX (aliquot)–Forte
(50 μl reactions)
P561F-50 2X 96 tubes ₩ 200,000 추가
4 2X TOPsimple™ DyeMIX (aliquot)–Forte
(50 μl reactions)
P562F-50 2X 960 tubes ₩ 1,600,000 추가
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Product description

This product is 2X TOPsimple™ DyeMIX-Forte that is aliquoted into PCR tube strips. For example, 8 strips of 12 tubes are provided for 96 PCR reactions (Cat. P561F). Since it contains loading dyes, the reaction mixtures after PCR are ready for agarose gel electrophoresis. This product includes nPfu-Forte (Cat. P410, P425), and is optimized to amplify high-fidelity DNA and DNA up to 10 kb long.

 

Characteristics

- Single-tube convenience

- Well-suited to analysis of a large number of samples.

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

- Blunt end product

 

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 (2X)

nPfu-Forte DNA polymerase: 0.2 unit/μl

nPfu-Forte buffer (containing 4 mM MgSO4)

- dNTP mixture: 0.4 mM each

- Stabilizer

- Dye: 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

table up to 18 months at -20 or 3 months at 4 n(Storage at -20 is recommended).

 

Standard reaction conditions

- PCR mixturea

Component

Reaction volume

(20 μl)

Reaction volume

(50 μl)

2X TOPsimple™ DyeMIX(aliquot)-Forte

1 tube

1 tube

Template DNAb (0.1~500 ng/μl)

1 μl

2.5 μl

Primer 1 (5 pmole/μl)

1 μl

2.5 μl

Primer 2 (5 pmole/μl)

1 μl

2.5 μl

Sterile water

up to 20 μl

up to 50 μl

aAssmeble the reaction mixture on ice

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

 

- PCR cycles

Step

Temperature

Time

Cycle

Initial denaturation

95

10 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.

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