Product description

2X TOPsimple™ DyeMIX-HOT is similarly formulated to the 2X TOPsimple™ PreMIX-HOT 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-HOT (Cat. P725, P750), and is optimized to increase the specificity and efficiency of target DNA amplification.

Characteristics

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

- 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 room temperature

- Colony PCR

Components (2X)

- nTaq-HOT DNA polymerase: 0.2 unit/μl

- nTaq-HOT 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 mixture

^aPlasmid DNA, 0.1 ng-30 ng; genomic DNA, 50 ng-500 ng

- PCR cycles

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.

●  Enzynomics_P510H_2X TOPsimple™ DyeMIX-HOT_manual

● Material Safety Data Sheet

- Enzynomics_MSDS_P510H_2X TOPsimple™ DyeMIX–HOT