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2X TOPsimple™ DyeMIX–HOT
No | Product Name | Cat.# | Conc | Size | Price(₩) | Add |
---|---|---|---|---|---|---|
1 | 2X TOPsimple™ DyeMIX–HOT | P510H | 2X | 1 ml | ₩ 50,000 |
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2 | 2X TOPsimple™ DyeMIX–HOT | P525H | 2X | 2.5 ml | ₩ 100,000 |
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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
2X
TOPsimple™ DyeMIX-HOT |
1 tube |
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.
● Material Safety Data Sheet