2×Universal Blue SYBR Green qPCR Master Mix With 40×Yellow Template Dilution Buffer 1mL
- Stock: In Stock
- Model: 0466
Product Description/Introduction
2×Universal Blue SYBR Green qPCR Master Mix is a specialized 2× premix for qPCR reactions using the SYBR Green I chimeric fluorescence method. The core component Taq DNA Polymerase is a hot-start DNA polymerase based on the antibody method of confinement, which can effectively inhibit non-specific amplification under low-temperature conditions, and at the same time with the reaction Buffer optimized for qPCR, it is very suitable for the qPCR reaction with high specificity and sensitivity, and it can obtain a good standard curve within a wide quantitative region, and quantify the target gene with accurate, reproducible and highly reliable. At the same time, the product contains a special ROX Passive Reference Dye, which is suitable for use in all qPCR instruments, and there is no need to adjust the ROX concentration on different instruments. Pre-mixed solution is added with blue dye, with the addition of tracer, while supporting the provision of yellow template diluent, when the template is diluted with yellow template diluent and added to the blue reaction pre-mixed solution, the color changes from blue to green, the reaction system can be formulated to play a role in tracing the reaction system process, to prevent the leakage of the addition or wrong addition. The spectra of this blue and yellow dye do not overlap with the qPCR dye and do not affect the results of the reaction.
Storage and Shipping Conditions
Ship with wet ice; store at -20℃, valid for 12 months.
Assay Protocol / Procedures
1. (Optional) Template Dilution
The Yellow Sample Buffer is supplied at a 40X concentration for DNA template dilution, which can accurately determine whether the template has been added to the qPCR reaction solution by the change of liquid color. Taking the 20-μL qPCR reaction system as an example, according to the dilution template amount added into the 20μL qPCR reaction solution, the corresponding dilution method of the original template can be referred to the following table (taking the original template diluted to 100μL as an example):
For example: 2μL diluted template should be added to 20-μL qPCR reaction system. Taking the 100μL Template Dilution system as an example. If the original template volume is 20μL, 25μL 40x Yellow Template Dilution Buffer is added, and then Nuclease-free Water 55μL is added to the total volume of 100μL. The 40x YELLOW Template Dilution Buffer is now 10x. Add 2μL of the diluted template into a 20-μL qPCR reaction system, and the final concentration of 40× YELLOW Template Dilution Buffer is 1x. Or if you do not want to use the template dilution for G3326-2, you can ignore this step.
1. 2. Recommended qPCR reaction system:
Component | 20 μL rxn | 50 μL rxn | Final Concentraction |
2×Universal Blue SYBR Green qPCR Master Mix | 10 μL | 25 μL | 1× |
0.4 μL | 1 μL | 0.2 μM | |
Reverse Primer (10μM)a | 0.4 μL | 1 μL | 0.2 μM |
Templateb | Variable | Variable | as required |
Nuclease-Free Water | Add to 20 μL | Add to 50 μL |
a: Usually, a good amplification effect can be obtained with the final concentration of 0.2 μM. When the reaction performance is poor, the primer concentration can be adjusted in the range of 0.2-1.0 μM.
b: The amount of template added varies depending on the number of copies of the target gene, and the appropriate amount of template addition is studied by gradient dilution. The best addition amount of template DNA in the 20 μL reaction system was less than 100 ng. When the cDNA (RT reaction solution) of RT-PCR reaction was used as template, the addition amount should not exceed 10% of the final qPCR volume.
2. 3. PCR reaction program (can be adjusted appropriately according to the instruments):
1-250x250.jpeg "2 × Fast SYBR Green qPCR Master Mix (High ROX) 1mL")
1-250x250.jpeg "2 × Fast SYBR Green qPCR Master Mix (Low ROX) 1mL")
