2×Super Kfx MasterMix (Dye)

Item Number

S665699

• Storage Temp: Store at -20°C
• Shipped In: Ice chest + Ice pads

Product Description

Product content

S665699	Component	1 mL	5 mL	Storage
S665699A	2×Super Kfx MasterMix (Dye)	1 mL	5×1 mL	-20℃. Avoid freeze/thaw cycle.
S665699B	ddH2O	1 mL	5×1 mL	-20℃. Avoid freeze/thaw cycle.

Product Introduction

This product is a premixed system composed of Super Kfx DNA Polymerase, Mg2+, dNTPs, PCR stabilizers and enhancers, with a concentration of 2 ×. Super Kfx DNA Polymerase is a fast and highly efficient high fidelity DNA polymerase with 5 ′ -3 ′ DNA polymerase activity and 3 ′ -5 ′ exonuclease activity. It has advantages such as strong amplification ability, high fidelity, and strong specificity. The addition of unique amplification enhancers and elongation factors in 2xMix results in a unique formula that makes the entire reaction system very stable, easy to operate, and suitable for amplification of various fragments and templates. This product is suitable for gene cloning, second-generation library amplification, gene directed mutagenesis, SNP amplification experiments, etc. This product has been added with dye (blue), and after the reaction is completed, agarose electrophoresis detection can be directly performed, making the operation convenient and simple.

Quality control

After testing, there is no exogenous nuclease activity and no residual host DNA, which can effectively amplify various templates.

Usage

The following are examples of conventional PCR reaction systems and reaction conditions. In practical operation, corresponding improvements and optimizations should be made based on different templates, primer structures, and target fragment sizes.

1. PCR reaction system

All operations should be carried out on ice. After each group is decomposed and frozen, please mix thoroughly. After use, please put it back at -20 ℃ for storage in a timely manner.

 

2. PCR reaction conditions

 

Attention:

1) Priority should be given to using the three-step method for amplification. If the three-step method cannot amplify the target product or the Tm value of the primer is greater than 68 ° C, please try the two-step method.

2) Denaturation: Pre denaturation of simple templates at 98 ℃ for 30 seconds to 1 minute. For complex templates, the pre denaturation time can be extended to 3 minutes.

3) Annealing: In general experiments, the annealing temperature is 3-5 ℃ lower than the Tm value of the primer. If the ideal amplification efficiency cannot be achieved, the annealing temperature should be gradually changed for optimization; When non-specific reactions occur, increase the annealing temperature appropriately.

4) Extension: The extension time should be set based on the length of the amplified fragment and the complexity of the template. The amplification efficiency of this product is 4-6 kb/min. For long fragments and high complexity templates, it is recommended to set 2-4 kb/min.

5) Cycle count: The number of cycles can be set based on the downstream application of the amplification product. If there are too few cycles, insufficient amplification, or too many cycles, the probability of mismatch will increase. Therefore, while ensuring product yield, the number of cycles should be minimized as much as possible.