Product Introduction
The One-step Probe RT-qPCR Kit V3 is designed for quantitative PCR detection using RNA as a template, such as viral RNA. Using gene-specific primers (GSP), reverse transcription and qPCR reactions are performed in a single tube, requiring no additional tube opening/pipetting, significantly increasing detection throughput and reducing the risk of contamination. This Kit introduced dUTP/UDG system. Heat-labile UDG can rapidly degrade the pollutants containing U at room temperature. When reverse transcription is performed at 55°C, heat-labile UDG will be inactivated rapidly without affecting the efficiency and sensitivity of RT-qPCR. Integrating the superior performance of Reverse Transcriptase and hot-start Taq DNA Polymerase, with the optimized buffer system, the detection sensitivity of One-step Probe RT-qPCR Kit can reach 0.1 pg total RNA or <10 copies of RNA template. The kit is available as a convenient Master Mix. 5 × One-step U+ Mix contains an optimized buffer system and dNTP/dUTP Mix, which is suitable for high-specificity, low template concentration and multiple detection systems with fluorescent labeled probes such as TaqMan®. This product is perfectly compatible with common quantitative PCR instruments, such as ABI, Roche, Bio-Rad, etc.
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
• Fast start-up, 30s hot start
• Reverse transcription at 55 °C to accommodate complex RNA templates
• Contains heat-labile UDG to reduce false-positive contamination
• High sensitivity for detection of low-abundance genes
• High stability, unchanged performance at 4°C and 37°C for one week
• The reaction system can be formulated up to 30 minutes in advance without changing performance
• Saves time by supporting fast program
Experimental Cases
Using V5011 for nucleic acid detection of coronavirus can accurately detect the E gene, N1 gene, N2 gene, and ORF1ab sequence
Components
Component | V5011 (200 rxns) | V5011-2 (1000 rxns) |
5X One-step U+ Mix | 800 μl | 1 ml × 4 |
One-step U+ Enzyme Mix V3 | 200 μl | 1 ml |
RNase-free ddH2O | 1 ml | 15 ml |
Storage Condition
Store at -20°C