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Polymerases

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2X TOPsimple™ DyeMIX (aliquot)–Tenuto

No Product Name Cat.# Conc Size Price(₩) Add
1 2X TOPsimple™ DyeMIX (aliquot)–Tenuto
(20 μl reactions)
P561 2X 96 tubes ₩ 70,000 추가
2 2X TOPsimple™ DyeMIX (aliquot)–Tenuto
(20 μl reactions)
P562 2X 960 tubes ₩ 560,000 추가
3 2X TOPsimple™ DyeMIX (aliquot)–Tenuto
(50 μl reactions)
P561-50 2X 96 tubes ₩ 140,000 추가
4 2X TOPsimple™ DyeMIX (aliquot)–Tenuto
(50 μl reactions)
P562-50 2X 960 tubes ₩ 1,120,000 추가
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Product description

This product is 2X TOPsimple™ DyeMIX-Tenuto that is aliquoted into PCR tube strips. For example, 8 strips of 12 tubes are provided for 96 PCR reactions (Cat. P561). Since it contains loading dye, the reaction mixtures after PCR are ready for agarose gel electrophoresis. This product includes nTaq-Tenuto (Cat. P225, P250), and is optimized to amplify DNA longer than 10 kb, which is difficult with common Taq polymearases. Thus, this product is improved in both fidelity (> 2 fold) of PCR product and amplification efficiency of longer PCR products.  

 

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

- Amplification of long DNA fragments (<10 kb)

- Amplification of high-complexity template DNA

- Primer extension

- Colony PCR

 

Components (2X)

nTaq-Tenuto: 0.2 unit/μl

nTaq-Tenuto 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 mixturea

Component

Reaction volume

(20 μl)

Reaction volume

(50 μl)

2X TOPsimple™ DyeMIX(aliquot)-Tenuto

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

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.

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