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Polymerases

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2X TOPsimple™ PreMIX-HOT

No Product Name Cat.# Conc Size Price(₩) Add
1 2X TOPsimple™ PreMIX-HOT P610H 2X 1 ml ₩ 50,000 추가
2 2X TOPsimple™ PreMIX-HOT P625H 2X 2.5 ml ₩ 100,000 추가
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Product description

2X TOPsimple™ PreMIX-HOT is a ready-to-use PCR mixture containing 2X concentration of DNA polymerase, reaction buffer, and dNTP. This product includes nTaq-HOT, and is optimized to increase the specificity and efficiency of target DNA amplification.

 

Characteristics

- 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 the room 

  temperature

- Colony PCR

- Labeling of DNA fragments with radioactive-isotopes

- Nucleotide sequencing 

 

Components (2X)

nTaq-HOT DNA polymerase: 0.2 unit/µl

nTaq-HOT buffer (containing 4 mM MgCl2)

- dNTP mixture: 0.4 mM each

- Stabilizer

 

Storage

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

 

Standard reaction conditions

- PCR mixture

2X TOPsimple™ PreMIX-HOT

10 μl

Template DNAa (0.1~500 ng/μl)

1 μl

Primer 1 (5 pmole/μl)

1 μl

Primer 2 (5 pmole/μl)

1 μl

Sterile water

up to 20 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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