詳細(xì)說(shuō)明
Lolina A/SAddress: Sindalsvej 30 8240 Risskov Danmark
Email: Info@lolina.dk
Website: https://lolina.dk
Product Specification
Product name | 2 × Lolina? AdvanceFast PCR Master Mix-With Dye |
Cat.No. | NaM201001-2 |
Size | 250 μL/1 mL/5×1 mL |
Storage and shipping | The product is shipped with dryice and can be stored at -25℃ ~ - 15 ℃ for 1 year. |
Product description
2×Lolina? AdvanceFast PCR Master Mix (With Dye) is a ready-to-use 2× pre-mixed solution containing Lolina? AdvanceFast High-Fidelity DNA Polymerase, dNTPs, and an optimized buffer system, which contains preadded electrophoresis indicators. The pre-mix contains pre- added electrophoresis indicator, PCR products can be directly electrophoresed, the amplification products are flat ends. 2×Lolina? AdvanceFast PCR Master Mix (With Dye) has the advantages of quick and easy, high sensitivity, high specificity, good stability, etc., the reaction system can be added with only the primers and templates. In addition, the product also contains a specific protective agent, so that the premix can still maintain stable activity after repeated freezing and thawing.
Instructions
1. Recommended PCR Reaction System
Components | Volume μL | Final concentration |
2 × Lolina? AdvanceFast PCR Master Mix (With Dye)a) | 25 | 1 × |
Templateb) | 0 | - |
Forward Primer (10 μmol/L)c) | 2 | 0.4 μmol/L |
Reverse Primer (10 μmol/L) | 2 | 0.4 μmol/L |
ddH20 | Up to 50 | - |
[Note]:
a) In 1× premixes containing 2 mM Mg2+ and 200 μM dNTPs.
b) Recommended range 10-200 ng, cDNA sample upload volume range not more than 1/10 of the reaction system, recommended 1-2.5 μL.
c) The final primer concentration in the PCR reaction system ranges from 0.2-1 μM, and 0.4 μM is recommended.
2.Reaction program
Cycle step | Temp. | Time | Cycles |
Initial denaturation | 98 °C | 30 sec | 1 |
Denaturation | 98 °C | 10 sec |
30-35 |
Annealinga) | 60 °C | 5 sec | |
Extensionb) | 72 °C | 5-10 sec/kb | |
Final extension | 72 °C | 2 min | 1 |
[Note]:
a ）Recommended temperature: 60°C, a temperature gradient can be set up to find the optimal temperature for primer annealing. The recommended annealing time is set to 5 sec and can be adjusted from 5-30 sec. Too long annealing time may result in diffuse amplification products on the gel.
b) Extension time: Recommended 5 sec/kb, can also be extended to 10 sec/kb as needed.
Notes
1. This product is for research use only.
2. Please operate with lab coats and disposable gloves for your safety.
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