Description
Product description
The kit is a ready-to-use PCR premixed solution containing HotStart Taq DNA Polymerase, dNTPs, and an optimized buffering system. Simply add primers and template for amplification, which greatly simplifies the experimental procedure and allows for high-throughput operations, enhancing the reproducibility of results. The hotStart Taq DNA Polymerase is a ligand-modified thermostable Taq DNA Polymerase, with a ligand that modulates the activity of the DNA polymerase in response to temperature changes. The enzyme activity is completely sealed at room temperature and is only released after heating at 95℃. The activation time for HotStart DNA Polymerase requires only 2-3 min and is compatible with existing PCR protocols. This product prevents non-specific amplification during sample preparation and reaction heating stages, effectively enabling genotyping experiments.
Components
|
Name
|
N132004E
|
N132004S
|
N132004M
|
N132004L
|
|
2×HotStart Genotyping PCR Master Mix
(With Dye)
|
1 mL
|
5×1 mL
|
50×1 mL
|
100×1 mL
|
Specifications
|
Concentration
|
2×
|
|
Hot Start
|
Built-in Hot Start
|
|
Overhang
|
3 '-A
|
|
Polymerase
|
Taq DNA Polymerase
|
|
Reaction speed
|
Standard
|
|
Product Type
|
PCR Master Mix (2x)
|
Shipping and Storage
Dry ice shipping. -15℃ ~ -25℃ storage, valid for two years.
Applications
This product is primarily applied to the genotyping of mice.
Notes
1. For your safety and health, please wear lab coats and disposable gloves for operation.
2. This product is for research use ONLY!This product is for research use ONLY!
Instructions
1. Reaction System
Table 1 Reaction system
|
Components
|
Size (μL)
|
Final concentration
|
|
2×HotStart Genotyping PCR Master Mix (With Dye)
|
25
|
1×
|
|
Template DNA
|
suitable
|
-
|
|
Forward primer (10 μmol/L)
|
2
|
0.4 μM
|
|
Reverse primer (10 μmol/L）
|
2
|
0.4 μM
|
|
ddH2O
|
to 50
|
-
|
Recommended usage of the different templates:
Table 2 Recommended usage of different templates
|
Type of template
|
Segment usage range (50 μL reaction system)
|
|
Genomic DNA
|
50–100 ng
|
|
Plasmid DNA
|
0.1-20 ng
|
|
cDNA
|
1-5 μL (Do not exceed 1/10 of the reaction system)
|
2. Amplification Protocol
Table 3 Amplification protocol
|
Cycle steps
|
Temperature (°C)
|
Time
|
Cycles
|
|
Predenaturation
|
95
|
5 min
|
1
|
|
Denaturation
|
95
|
30 sec
|
35
|
|
Annealing
|
50-60
|
30 sec
|
|
Extension
|
72
|
30-60 sec/kb
|
|
Final extension
|
72
|
10 min
|
1
|
【Note】:
a. *Recommended Annealing Temperature and Time: The annealing temperature is recommended to be set at 50-60℃. The annealing time is recommended to be set at 30 sec, with adjustments possible within the range of 20-30 sec. Depending on the need, a temperature gradient can be established to explore the optimal annealing temperature and time for primer annealing.
b. **Extension Temperature and Time: The temperature is recommended to be 72℃. The time is recommended to be 30-60 sec/kb.
c. **PCR Amplification Products: Please store the PCR amplification products at -20℃ to prevent DNA degradation.
