Product Introduction
The DSPathTM 4X One-Step Multiplex Master Mix (High ROX+) is used to perform one-step multiplex real-time PCR applications with any gene-specific primer and probe sets in quantitative instruments that require high concentrations of ROX, and is suitable for both RNA and DNA targets. This master mix is formulated with optimized buffer components to accommodate multiplex amplification of up to four RNA or DNA target sequences in a single reaction. The master mix is supplied at a 4X concentration that allows to input more sample into each reaction, increasing sensitivity even in low-volume reactions.
We offer 4 formulations of this master mix, which have different buffers to meet amplification requirements of different primers:
Product Name | Cat. No. | Spec. | Quantity |
DSPathTM 4X One-Step Multiplex Master Mix | V5005 V5006 | 200 rxns 5000 rxns | 1 mL × 1 25 mL × 1 |
DSPathTM 4X One-Step Multiplex Master MixB | V5005B V5006B | 200 rxns 5000 rxns | 1 mL × 1 25 mL × 1 |
DSPathTM 4X One-Step Multiplex Master MixC | V5005C V5006C | 200 rxns 5000 rxns | 1 mL × 1 25 mL × 1 |
DSPathTM 4X One-Step Multiplex Master MixD | V5005D V5006D | 200 rxns 5000 rxns | 1 mL × 1 25 mL × 1 |
Easy to use
Simply add primers, probes, and RNA/DNA templates for rapid reactions.
Figure 1. One-Step RT-qPCR workflow using DSPathTM 4X One-Step Multiplex Master Mix
Selection Guide of One-Step Kit (probe based):
Cat. No. | V5001/V5002 | V5005/V5006 | V5005L/V5006L | V5007/V5008 | V5007L/V5008L | V5009/V5010 | V5011 | |
Product Name | One-step Probe RT-qPCR Kit | DSPathTM 4X One-Step Multiplex Master Mix | DSPathTM 4X One-Step Multiplex Master Mix (Lyophilized) | 2X One Step Prime RT-qPCR Mix | 2X One Step Prime RT-qPCR Mix (Lyophilized) | One-step Probe RT-qPCR Kit V2 | One-step Probe RT-qPCR Kit V3 | |
Mixture Format | Enzyme Mix+2X Buffer Mix | 4X All-in Mix | 4X All-in Lyophilized Powder | 2X All-in Mix | 2X All-in Lyophilized Powder | Enzyme Mix+2X Buffer Mix | Enzyme Mix+5X Buffer Mix | |
Mode/Time | S: ~120 min | F: ~60 min S: ~120 min | F: ~60 min S: ~120 min | F: ~60 min S: ~120 min | F: ~60 min S: ~120 min | F: ~60 min S: ~80 min | F: ~45 min S: ~80 min | |
RT TEMP | 50~55°C | 48~55°C | 48~55°C | 48~55°C | 48~55°C | 50~55°C | 50~55°C | |
PCR Enzyme | Antibody-modified hotstart Taq | Antibody-modified hotstart Taq | Antibody-modified hotstart Taq | Antibody-modified hotstart Taq | Antibody-modified hotstart Taq | Antibody-modified hotstart Taq | Antibody-modified hotstart Taq | |
Heat-labile UDG | + | + | + | + | + | + | + | |
Sensitivity | Very high | Very high | Very high | Very high | Very high | Ultra high | Ultra high |
S: standard mode; F: fast mode
V5005, V5009, and V5011 are recommended
Application
• Probe gene expression analysis
• Probe Low-copy gene detection
• Probe microarray validation
• Probe gene knockdown validation
Features
• Robust one-step, all-in-one master mix system for easy reaction assembly
• Detect multiple targets in one reaction
• High sensitivity to detect low-copy targets
• Tolerance of inhibitors commonly found in clinical samples
• Eliminates the risk of cross contamination associated with two-step RT-qPCR protocols
• Premixed with high concentration of ROX reference dye
Experimental Cases
1. Comparable to first-line brands
Compare with 6 different brands of similar products to detect samples containing the novel coronavirus at low or high concentrations, #V5005 has excellent detection rate and accuracy.
Figure 2. detection of low SARS-CoV-2 sample
Figure 3. detection of high SARS-CoV-2 sample
Stable performance
DSPath 4X One-Step Multiplex Master Mix is very stable, and you can get the benefit from high stability: keep the master mix in the refrigerator for up to 4 weeks and profit from a quick setup without thawing first.
The stability test of DSPath 4X One-Step Multiplex Master Mix/MixB/MixC/MixD was performed at 37°C for 8 days by detecting four targets of SARS-CoV-2. The amplification efficiency of the four products did not change significantly, which means all of them could maintain high stability, and are easy to be transported and stored for a long time.
Figure 4. amplification curve of stability test of DSPathTM 4X One-Step Multiplex Master Mix/MixB/MixC/MixD
Figure 5. Ct value of stability test of DSPathTM 4X One-Step Multiplex Master Mix/MixB/MixC/MixD
Components
Component | V5005H (200 rxns) | V5006H (5000 rxns) |
DSPath 4X Multiplex Master Mix (High ROX+) | 1 ml × 1 | 25 ml × 1 |
Storage Condition
Store at -20°C