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2X TOPsimple™ PreMIX–Tenuto

No Product Name Cat.# Conc Size Price(₩) Add
1 2X TOPsimple™ PreMIX–Tenuto P610 2X 1 ml ₩ 55,000 추가
2 2X TOPsimple™ PreMIX–Tenuto P625 2X 2.5 ml ₩ 110,000 추가
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Product description

2X TOPsimple™ PreMIX-Tenuto is a ready-to-use PCR mixture containing 2X concentration of DNA polymerase, reaction buffer, and dNTP. 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

- 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

- Labeling of DNA fragments with radioactive-isotopes

- Nucleotide sequencing

 

Components (2X)

nTaq-Tenuto: 0.2 unit/µl

nTaq-Tenuto 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 mixturea

2X TOPsimple™ PreMIX-Tenuto

10 μl

Template DNAb (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

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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