Product description

2× Hieff Canace™ AdvanceFast PCR Master Mix (With Dye) is a ready-to-use 2× pre-mixed solution containing Hieff Canace™ AdvanceFast High-Fidelity DNA Polymerase, dNTPs, and an optimized buffer system, which contains pre-added electrophoresis indicators. The pre-mix contains pre-added electrophoresis indicator, PCR products can be directly electrophoresed, the amplification products are flat ends. 2× Hieff Canace™ 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.

Ultra-high fidelity: Low mismatch rate in mutation-prone high GC gene;
Fast speed: The speed is as fast as 5 sec/kb;
Wide applicability: It can be used to amplify genes with 20-80% GC content, and tolerate 20% blood and mouse lysates;
Good stability: Stored at 37°C for 7 days without affecting performance;
Easy use: Just adding primers and templates for amplification, with blue loading dye for direct electrophoresis.

Specifications

Cat.No.	10164ES01	10164ES03	10164ES08
Size	250 μL	1 mL	5×1 mL

Figures

Figure: Hieff Canace™ AdvanceFast High-Fidelity DNA Polymerase has high fidelity compared to other brands' products (A). The assay was done by examining the mutation frequency of PCR products of 6 GC-rich (about 80%) segments amplified from the genomes of different species. The PCR products amplified by Hieff Canace™ AdvanceFast High-Fidelity DNA Polymerase were cloned into vectors, and 100 selected single clones for each sequence were subjected to Sanger sequencing.

Hieff Canace™ AdvanceFast High-Fidelity DNA Polymerase has high amplification speed (B), high GC amplification compatibility (C), and blood tolerance (D). Results from customers' PCR and sequencing show that Hieff Canace™ AdvanceFast High-Fidelity DNA Polymerase has good performance in PCR yields and specificity (E-H).

 

Storage

This product should be stored at -25~-15℃ for 1 years.

Instructions 

1. Recommended PCR reaction systems.

Components	Volume（μL）	Final concentration
2× Hieff Canace™ AdvanceFast PCR Master Mix (With Dye)*	25	1×
Template**	x	-
Forward Primer（10 μmol/L）***	2	0.4 μmol/L
Reverse Primer（10 μmol/L）	2	0.4 μmol/L
ddH2O	Up to 50	-

Table 1 PCR reaction system

*In 1× premixes containing 2 mM Mg2+ and 200 μM dNTPs.

**Recommended range 10-200 ng, cDNA sample upload volume range not more than 1/10 of the reaction system, recommended 1-2.5 μL.

***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℃	30 sec	1
Denaturation	98℃	10 sec	30-35
Annealing*	60℃	5 sec
Extension**	72℃	5-10 sec/kb
Final extension	72℃	2 min	1

Table 2 PCR reaction program

*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.

**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.