컨텐츠 바로가기 영역
주메뉴로 바로가기
본문으로 바로가기

All products

Home > PRODUCTS > All products

2X TOPsimple™ DyeMIX–Tenuto

No Product Name Cat.# Conc Size Price(₩) Add
1 2X TOPsimple™ DyeMIX–Tenuto P510 2X 1 ml ₩ 55,000 추가
2 2X TOPsimple™ DyeMIX–Tenuto P525 2X 2.5 ml ₩ 110,000 추가
Product Name Cat.# Size Price(₩) EA Total(₩) 삭제
Total : 장바구니에 담기 바로 구매 하기
plus minus 삭제

Product description

2X TOPsimple™ DyeMIX-Tenuto is similarly formulated to the 2X TOPsimple™ PreMIX-Tenuto except that it contains loading dyes for further convenience of use. Thus, the reaction mixtures after PCR cycles 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 products and amplification efficiency of longer PCR products.

 

Characteristics

- 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 (>5-15 kb)

- Amplification of high-complexity template DNA such as cDNA and genomic 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

2X TOPsimple™ DyeMIX-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.


List