Product Introduction
The GDSlyo One-step Probe RT-qPCR Kit is a glycerol-free one-step RT-qPCR reagent containing lyophilized excipients for the detection of RNA templates that can be lyophilized with a suitable lyophilization process. The difficulties in the development of lyophilized products include three aspects:1. The removal of liquid reagents affects the lyophilized components, and the stability is poor; 2. It is difficult to freeze-dry and mold, and the yield rate is low; 3. Poor long-term storage stability of lyophilized finished products. Through the optimization of functional proteins, the matching and screening of excipients and lyophilization processes, this product successfully solves the above three problems. Through the screening of enzymes and excipients with high stability, the storage stability of this product is consistent with that of glycerin-containing products; By selecting the appropriate lyophilized excipients and matching the lyophilization process, the yield and stability of the lyophilized products are improved, so that the lyophilized products can be stored at room temperature for more than one year. In addition, this product offers the advantages of conventional glycerol reagents, including good multiplex amplification performance, high specificity, and high sensitivity. The dUTP/UDG anti-contamination system is incorporated into the reagents, which can be used at room temperature to eliminate the influence of amplification product contamination on qPCR results and ensure the accuracy of results.
Cat. No./Spec.
Cat. No. | V5013-A | V5013-B | V5013-C |
20-μl reaction Nos. | 200 rxns | 1,000 rxns | 5,000 rxns |
Components
Component | V5013-A | V5013-B | V5013-C |
5X GDSlyo One-step Buffer [1] | 800 μl | 4 ml | 20 ml |
GDSlyo One-step Enzyme Mix [2] | 200 μl | 1 ml | 5 ml |
3X Excipientc [3] | 1.2 ml | 6 ml | 30 ml |
50X ROX Reference Dye 1 [4] | 80 μl | 400 μl | 1 ml × 2 |
50X ROX Reference Dye 2 [4] | 80 μl | 400 μl | 1 ml × 2 |
RNase-free ddH2O | 1 ml × 3 | 15 ml | 75 ml |
[1] Contains dNTP/dUTP mix, Mg2+.
[2] Contains Reverse Transcriptase, RNase inhibitor, Heat-labile UDG and Taq DNA Polymerase.
[3] Lyophilized excipients, protect the activity of enzymes during the lyophilization process, maintain the morphology and long-term storage stability of lyophilized products.
[4] Used to correct the fluorescence signal error between holes. 50X ROX Reference Dye 1 is used for ABI 7900HT/ 7300 real-time PCR System and StepOne Plus; ABI 7500, 7500 Fast real-time PCR System, Stratagene Mx3000P using 50X ROX Reference Dye 2; Roche, bio-rad Real Time PCR Systems do not have to use ROX.
Storage
This reagent should be kept at -20°C.
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
• One-step RT-qPCR
• This kit is suitable for fluorescence quantification by probe method
• This kit is compatible with many real-time systems
• Hot-start technology brings high specificity and reproducible amplification
• dUTP/UDG system, effectively prevent PCR product contamination