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2X TOPsimple™ DyeMIX (aliquot)-multi HOT
(UDG plus)

No Product Name Cat.# Conc Size Price(₩) Add
1 2X TOPsimple™ DyeMIX (aliquot)-multi HOT
(UDG plus)
P561HCU 2X 96 tubes ₩ 100,000 추가
2 2X TOPsimple™ DyeMIX (aliquot)-multi HOT
(UDG plus)
P562HCU 2X 960 tubes ₩ 800,000 추가
Product Name Cat.# Size Price(₩) EA Total(₩) 삭제
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Product description

2X TOPsimple™DyeMIX-multi HOT (UDG plus) is aliquoted into PCR tube strips. For example, 8 strips of 12 tubes are provided for 96 PCR reactions (Cat. P561HCU). Since it contains loading dyes, the reaction mixtures after PCR are ready for agarose gel electrophoresis. This product can be used in general multiplex PCR and genotyping experiment related to genetic diagnostics because it produces up to 20 different amplified products within a single tube reaction. 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)

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

- Carry-over contamination control: contains UDG

 

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-multi HOT DNA polymerase: 0.2 unit/μl

- Uracil-DNA Glycosylase

- nTaq-multi HOT 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 months at -20 or 3 months at 4 (Storage at -20 is recommended).

 

Standard reaction conditions

- PCR mixturea

2X TOPsimple™ DyeMIX(aliquot)-multi HOT (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 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 mixture 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

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

 


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