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Polymerases

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2X TOPsimple™ DyeMIX (aliquot)–HOT

No Product Name Cat.# Conc Size Price(₩) Add
1 2X TOPsimple™ DyeMIX (aliquot)–HOT
(20 μl reactions)
P561H 2X 96 tubes ₩ 65,000 추가
2 2X TOPsimple™ DyeMIX (aliquot)–HOT
(20 μl reactions)
P562H 2X 960 tubes ₩ 520,000 추가
3 2X TOPsimple™ DyeMIX (aliquot)–HOT
(50 μl reactions)
P561H-50 2X 96 tubes ₩ 130,000 추가
4 2X TOPsimple™ DyeMIX (aliquot)–HOT
(50 μl reactions)
P562H-50 2X 960 tubes ₩ 1,040,000 추가
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Product description

This product is 2X TOPsimple™ DyeMIX-HOT that is aliquoted into PCR tube strips. For example, 8 strips of 12 tubes are provided for 96 PCR reactions (Cat. P561H). Since it contains loading dyes, the reaction mixtures after PCR are ready for agarose gel electrophoresis. This product includes nTaq-HOT (Cat. P725, P750), and is optimized to increase the specificity and efficiency of target DNA amplification.

 

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)

- A-tail formation at 3’ ends of amplified duplex DNA

 

Applications

- High specific amplification of DNA fragments shorter than 3 kb.

- Amplification of cDNA and genomic DNA.

- Amplification of template DNA with secondary or higher ordered structure that is resistant to PCR amplification

- Primer extension

- Multiplex PCR

- Well-suited for an automated PCR machines, for which PCR reaction mixtures are prepared at room temperature

- Colony PCR

 

Components (2X)

nTaq-HOT DNA polymerase: 0.2 unit/μl

nTaq-HOT buffer (containing 4 mM MgCl2)

- dNTP mixture: 0.4 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 18 months at -20 or 3 months at 4 (Storage at -20 is recommended).

 

Standard reaction conditions

- PCR mixture

Component

Reaction volume

(20 μl)

Reaction volume

(50 μl)

2X TOPsimple™ DyeMIX(aliquot)-HOT

1 tube

1 tube

Template DNAa (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

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

  

- PCR cycles

Step

Temperature

Time

Cycle

Initial denaturationa

95

10 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 at 4; may add 10 mM EDTA until use to prevent DNA degradation

aAt least 10 min of initial denaturation time is required to fully activate the chemically modified PCR DNA polymerase

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

 

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