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2X TOPsimple™ DyeMIX (aliquot)-multi HOT
| No | Product Name | Cat.# | Conc | Size | Price(￦) | Add |
|---|---|---|---|---|---|---|
| 1 | 2X TOPsimple™ DyeMIX (aliquot)-multi HOT (20 μl reactions) |
P561HC | 2X | 96 tubes | ￦ 90,000 |
|
| 2 | 2X TOPsimple™ DyeMIX (aliquot)-multi HOT (20 μl reactions) |
P562HC | 2X | 960 tubes | ￦ 720,000 |
|
| 3 | 2X TOPsimple™ DyeMIX (aliquot)-multi HOT (50 μl reactions) |
P561HC-50 | 2X | 96 tubes | ￦ 180,000 |
|
| 4 | 2X TOPsimple™ DyeMIX (aliquot)-multi HOT (50 μl reactions) |
P562HC-50 | 2X | 960 tubes | ￦ 1,440,000 |
|
Product description
2X TOPsimple™DyeMIX-multi HOT is aliquoted
into PCR tube strips. For example, 8 strips of 12 tubes are provided for 96 PCR
reactions (Cat. P561HC). Since it contains loading dyes, the reaction mixtures
after PCR are ready for agarose gel electrophoresis. This product can be
used in general multiplex PCR and genotyping experiment related to genetic
diagnostics because it produces up to 20 different amplified products within a
single tube reaction.
Characteristics
- Single-tube convenience
- Well-suited to analysis of a large number of samples.
- 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-multi HOT DNA Polymerase:
0.2 unit/μl
- nTaq-multi 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 mixturea
|
Component |
Reaction volume (20 μl) |
Reaction volume (50 μl) |
|
2X TOPsimple™ DyeMIX(aliquot)-multi HOT |
1 tube |
1 tube |
|
Template DNAb (0.1~500 ng/μl) |
1 μl |
2.5 μl |
|
Primer 1 (5 pmole/μl) |
1 μl |
2.5 μl |
|
Primer 2 (5 pmole/μl) |
1 μl |
2.5 μl |
|
Sterile water |
up to 20 μl |
up to 50 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 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
- Enzynomics_MSDS_P561HC_2X TOPsimple™ DyeMIX (aliquot)-multi HOT
