2×Multiplex Probe qPCR Master Mix with UDG（None ROX）
- Stock: In Stock
- Model: 0438
Description/Introduction
The product is a 2×fluorescence quantitative PCR premixture (None ROX) based on probe method, it contains hot-start TaqDNA polymerase, dNTP and necessary buffer components, and can simultaneously perform up to quadruple fluorescence quantitative PCR reaction in the same reaction tube. The premix does not contain correction dyes and is suitable for equipment without ROX correction. The addition of blue non-fluorescent dye to this product can avoid sampling errors. In addition, the product also includes Uracil-DNAGlycosylase (UDG) and specially optimized dUTP addition amount, which can prevent template residual contamination and improve the specificity, sensitivity and accuracy of the product.
Storage and Handling Conditions
Shipped with wet ice and stored at -20°C; valid for up to12 months.
Product Contents
Component Number | Component | G3347-01 | ||
0438 | 2×Multiplex Probe qPCR Master Mix with UDG (None ROX) | 1 mL | ||
Manual | ||||
Before starting (please read carefully)
1. Real-time fluorescence quantitative PCR instrument is required.
2. Special qPCR reaction tubes or reaction plate for experiment are required but not supplied in this kit.
3. QPCR primers and probes (reference primers and probe design principles) are required but not supplied in this kit.
Assay Protocol / Procedures
1. Recommend PCR reaction system:
Component | 20 μL rxn | Final Concentration |
2×Multiplex Probe qPCR Master Mix with UDG (None ROX) | 10 μL | 1× |
Forward Primer (10 μM)a | Variable | 0.1-0.5 μM |
Reverse Primer (10 μM)a | Variable | 0.1-0.5 μM |
Probe (10 μM)a | Variable | 0.1-0.5 μM |
Templateb | Variable | as required |
Nuclease-free Water | Add to 20 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 addition varies with the number of copies of target genes in the template solution. Gradient dilute the template and investigate the appropriate amount of template addition. In the 20 μL reaction system, the amount of template DNA should be less than 100 ng. When using cDNA (RT reaction solution) of RT-PCR reaction as template, the addition amount should not exceed 10% of the total volume of PCR reaction solution.
2. PCR reaction procedure (can be adjusted according to the model):
A. Two-step method | B.Three-step method | |||||||||
Stage | Step | Cycle | Temp | Time | Stage | Step | Cycle | Temp | Time | |
Stage 1 | UDG incubation | 1 | 50°C | 2 min | Stage 1 |
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