【 Product Overview】
This product is a 2× concentration premixed reagent, containing Taq DNA Polymerase, Mg2+, dNTP and an optimized buffer solution. It is easy to operate and reduces the possibility of contamination, enabling high-throughput detection. Just add primers and templates to the reaction system to carry out the PCR reaction. The protective agent added to the system enables the 2×Taq PCR Mixture to maintain high activity after repeated freeze-thaw cycles. The product contains dyes, and the PCR products can be directly subjected to electrophoresis detection.
【 Product Features】
1. The product is a 2× premix liquid, which is simple and convenient to use.
The best effect is achieved when the amplification product is within 2kb.
3. Add an "A" base to the 3 'end of the amplification product, and after purification, it can be directly used for TA cloning.
4. The product contains dyes, and the PCR products can be directly subjected to electrophoresis detection.
【 Storage Conditions】
Store at a constant temperature of -20℃ for two years (transport at -20-8℃). Avoid repeated freezing and thawing. It can be stored for a short period at 4℃ for frequent use.
【 Experimental Procedure】
Preparation of the reaction system
| Reagent |
Reagent Volume(µL per sample) |
Final
Conc. |
| Mix A |
Mix B |
| Taq PCR Mix (2×) |
10 |
25 |
1× |
| Forward Primer (10 μM) |
0.5 |
1.0 |
0.25 μM |
| Reverse Primer (10 μM) |
0.5 |
1.0 |
0.25 μM |
| DNATemplate* |
Variable |
Variable |
/ |
| PCR Grade H2O |
To 20 |
To 50 |
/ |
| Total volume per reaction |
20 |
50 |
|
Note:
For different types of DNA samples, the amount of Template added varies. Taking a reaction volume of 20μL as an example, the recommended dosage of Template is as follows:
| DNA Template |
Reagent Volume |
| Genomic DNA |
20 ng ~ 100 ng |
| Plasmid DNA |
50 pg ~ 10 ng |
| cDNA (Product stock solution |
1 μL to 5 μL (not exceeding 1/10 of the reaction volume) |
- Reaction procedure
| Initial Denaturation |
95℃ |
2 min |
|
|
| Denaturation |
95℃ |
25 sec |
 |
35-40Cycle |
| Annealing*1 |
60℃ |
25 sec |
| Extension*2 |
72℃ |
1 min |
| Final Extension |
72℃ |
7 min |
|
|
Note:
The Annealing temperature is set according to the actual annealing temperature (Ta) of the primers.
For molecular identification, the extension speed is 30sec/kb; for gene cloning, it is recommended to use 60sec/kb.
3. Detection of amplification products
After PCR amplification is completed, agarose gel electrophoresis detection can be directly carried out.
【 Notes】
1. The product should be thawed on ice and repeated freezing and thawing should be avoided.
2. Before use, gently invert the reagent bottle up and down several times to ensure that all reagent components are thoroughly mixed. Unmixed reagents will have a reduced reaction performance.
3. The PCR reaction preparation process should be carried out on ice to reduce non-specific amplification generated during the reaction preparation stage.
4. Please read this instruction carefully and use this product strictly in accordance with the operation procedures and the recommended dosage.
5. This product is for scientific research use only and must not be used for clinical diagnosis.
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