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Polymerases

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2X TOPsimple™ DyeMIX (aliquot)-Tenuto (UDG plus)

No Product Name Cat.# Conc Size Price(₩) Add
1 2X TOPsimple™ DyeMIX (aliquot)-Tenuto
(UDG plus)
P561U 2X 96 tubes ₩ 80,000 추가
2 2X TOPsimple™ DyeMIX (aliquot)-Tenuto
(UDG plus)
P562U 2X 960 tubes ₩ 640,000 추가
Product Name Cat.# Size Price(₩) EA Total(₩) 삭제
Total : 장바구니에 담기 바로 구매 하기
plus minus 삭제

Product description

2X TOPsimple™DyeMIX(aliquot)-Tenuto (UDG plus) is aliquoted into PCR tube strips. For example, 8 strips of 12 tubes are provided for 96 PCR reactions (Cat. P561U). Since it contains loading dye, the reaction mixtures after PCR are ready for agarose gel electrophoresis. This product includes nTaq-Tenuto (UDG plus) (Cat. P225AU), 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. UDG and dUTP are included in the mixture to prevent the reamplification of carry-over PCR products between reactions.

 

Characteristics

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

- Carry-over contamination control: contains UDG

- 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 DNA polymerase: 0.2 unit/μl

- Uracil-DNA Glycosylase

nTaq-Tenuto buffer (containing 4 mM MgCl2)

- dNTP/dUTP 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 years at -20 or 3 months at 4 (Storage at -20 is recommended).

 

Standard reaction conditions

- PCR mixturea

2X TOPsimple™ DyeMIX(aliquot)-Tenuto (UDG plus)

1 tube (10 μl)

Template DNA(0.1~500 ng/μl)

μl

Primer 1 (5 pmole/μl)

μl

Primer 2 (5 pmole/μl)

μl

Sterile water

up to 20 μl

aAssemble the reaction mixture on ice

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

 

- PCR cycle

Step

Temperature

Time

Cycle

Pre-incubation (for UDG)

25

10 min

1

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

aRecommended annealing temperatures is 5 to 10 below the lower Tm of the two primers used.

 


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