Description
Product description
The kit is a kind of conventional PCR premixed solution which is ready to use, including Taq DNA Polymerase, dNTP mix, MgCl2 and optimized buffer. During the reaction, only the primer and template can be added for amplification, which greatly simplifies the operation steps of experiment. This product contains excellent stabilizers and can be stored for 3 months at 4℃. The PCR product have 3 '-dA protrusion and can be easily cloned into T vector.
Components
|
Components No.
|
N132003E
|
N132003S
|
N132003M
|
N132003L
|
|
Size
|
1 mL
|
5×1 mL
|
50×1 mL
|
100×1 mL
|
Specifications
|
Fidelity (vs. Taq)
|
1×
|
|
Hot Start
|
No
|
|
Overhang
|
3 '-A
|
|
Polymerase
|
Taq DNA Polymerase
|
|
Reaction Format
|
SuperMix or Master Mix
|
|
Reaction Speed
|
Standard
|
|
Product Type
|
PCR Master Mix (2x)
|
Shipping and Storage
Dry ice shipping. -15℃ ~ -25℃ storage, valid for two years.
Notes
1. PCR products with 2×PCR Master Mix are not suitable for polyacrylamide gel electrophoresis.
2. For your safety and health, please wear lab coats and disposable gloves for operation.
3. This product is for research use ONLY!
Instructions
1. Reaction System
Table 1 Reaction system
|
Components
|
Size (μL)
|
|
Template DNA
|
suitable
|
|
Primer 1 (10 μmol/L)
|
2
|
|
Primer 2 (10 μmol/L）
|
2
|
|
2×PCR Master Mix
|
25
|
|
ddH2O
|
to 50
|
【Note】:
1) Template usage: 50-200 ng genomic DNA; 0.1-10 ng plasmid DNA.
2) Mg2+ concentration: This product contains 3 mM of MgCl2, suitable for most PCR reactions.
2. Amplification Protocol
Table 2 Amplification protocol
|
Cycle steps
|
Temperature (°C)
|
Time
|
Cycles
|
|
Predenaturation
|
94
|
5 min
|
1
|
|
Denaturation
|
94
|
30 sec
|
35
|
|
Annealing
|
50-60
|
30 sec
|
|
Extension
|
72
|
30-60 sec/kb
|
|
Final extension
|
72
|
10 min
|
1
|
【Note】:
a. Annealing temperature: Please refer to the theoretical Tm value of primers. The annealing temperature can be set to 2-5℃ lower than the theoretical value of the primer.
b. Extention time: For molecular identification, 30 sec/kb is recommended. For gene cloning, 60 sec/kb is recommended.
Application example
Figure 1 The expected 1.2 kb PCR products can be amplified with 2× PCR Master Mix.
The Master Mix was stored at -20℃ for 1 year following another 3 months at 4℃ and 1 month at 25℃. Template: Arabidopsis genome. Annealing temperature: 60℃. Extension time: 40 sec.
