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2X TOPsimple™ DyeMIX–Tenuto
No | Product Name | Cat.# | Conc | Size | Price(₩) | Add |
---|---|---|---|---|---|---|
1 | 2X TOPsimple™ DyeMIX–Tenuto | P510 | 2X | 1 ml | ₩ 55,000 |
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2 | 2X TOPsimple™ DyeMIX–Tenuto | P525 | 2X | 2.5 ml | ₩ 110,000 |
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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.
● Material Safety Data Sheet