2xGoldStar MasterMix

Item Number

G665843

• Storage Temp: Store at -20°C,Avoid repeated freezing and thawing
• Shipped In: Ice chest + Ice pads

Product Description

2 x GoldStar MasterMix is a pre mixed system composed of GoldStar DNA Polymerase, PCR Buffer, Mg2+, dNTPs, PCR stabilizers and enhancers. The pre mixed PCR mixture makes the operation simpler and faster, minimizing human error and contamination. The GoldStar DNA Polymerase contained in this product is a chemically modified, novel and highly efficient Taq DNA Polymerase. The enzyme's activity is completely blocked at room temperature, making it inactive at low or room temperature, effectively avoiding non-specific amplification caused by non-specific binding of primers and templates or primer dimers at room temperature. The activation of the enzyme requires incubation at 95 ℃ for 10 minutes. The unique buffer system makes the application of enzymes more extensive, enabling efficient amplification of high GC content, complex secondary structures, and low copy templates. The unique MasterMix formula enhances the stability of the entire reaction system. Using this product for PCR amplification, the 3 'end of the PCR product contains an "A" base, which can be directly used for T/A cloning. This product does not contain dyes. After the PCR program is completed, an appropriate amount of sample buffer can be added as needed for electrophoresis operation. This product has strong specificity and does not require gel recovery to remove impurities after PCR amplification. It can be directly used for downstream cloning or chip hybridization experiments. Mainly used for routine PCR, RT-PCR, multiplex PCR, and gene chip detection, especially suitable for PCR reactions that require high specificity.

G665843	Component	5 mL	25 mL	Storage
G665843A	2×GoldStar MasterMix	5×1 mL	5×5 mL	-20℃. Avoid freeze/thaw cycle.
G665843B	ddH2O	5×1 mL	5×5 mL	-20℃. Avoid freeze/thaw cycle.

Notes: 2×GoldStar Taq MasterMix contains GoldStar DNA Polymerase, 3.4 mM MgCl2 and 400 µM each dNTP.

Quality control
No exogenous nuclease activity detected; PCR method for detecting host free residual DNA; Can effectively amplify single copy genes in the human genome; Store at 2-8 ° C for three months without significant changes in activity.
Usage
The following is an example of a PCR reaction system and reaction conditions for amplifying a 1 kb fragment using human genomic DNA as a template. In practical operation, corresponding improvements and optimizations should be made based on the different template primer structures and target fragment sizes.

1. PCR reaction system

Reagent	50 μl Reaction system	Final concentration
2×GoldStar MasterMix	25 μl	1 ×
Forward Primer，10 µM	2 μl	0.4 μM
Reverse Primer，10 µM	2 μl	0.4 μM
Template DNA	<0.5 μg	<0.5 μg/50 μl
ddH2O	up to 50 μl	/

Attention: The primer concentration should be between 0.1 and 1.0 as the final concentration μ M serves as a reference for setting the range. In the case of low amplification efficiency, the concentration of primers can be increased; When non-specific reactions occur, the primer concentration can be reduced to optimize the reaction system.
2. PCR reaction conditions

Step	Temperature	Time	/
Pre denaturation	95℃	10 min	/
Denaturation	95℃	30 s	30-40 cycles
Annealing	55-65℃	30 s	30-40 cycles
Extension	72℃	60 s	30-40 cycles
Final extension	72℃	5 min	/

Attention:
1) In general experiments, the annealing temperature is 5 ℃ lower than the melting temperature Tm of the amplification primer, and the annealing time is generally 30-60 seconds. If the ideal amplification efficiency cannot be achieved, the annealing temperature should be appropriately reduced; When non-specific reactions occur, increase the annealing temperature to optimize the reaction conditions.
2) The extension time should be set according to the size of the amplified fragment. The amplification efficiency of GoldStar DNA Polymerase contained in this product is 1-2 kb/min.
3) The number of cycles can be set based on the downstream application of the amplification product. If the number of cycles is too small, the amplification amount is insufficient; If there are too many cycles, the probability of mismatches will increase, and non-specific backgrounds will be severe. So, while ensuring product yield, the number of cycles should be minimized as much as possible.
4) This product must achieve enzyme activation under pre denaturation conditions of 95 ℃ and 10 minutes.