SYBR Green One-Step qRT-PCR Kit with Tracking Dyes
$212.00 - $852.00
$1,065.00
All products have special prices for bulk purchase, please contact for more details if required.
Cat. No.: SOQRPT-100 (for 100T)
Cat. No.: SOQRPT-500 (for 500T)
Description
SYBR Green One-Step qRT-PCR Kit with Tracking Dyes is a high-quality kit based on SYBR Green dye for one-step reverse transcription real-time quantitative PCR, also known as qRT-PCR (Quantitative Reverse Transcription PCR) or Real-time RT-PCR. It includes a dual-dye tracking system for highly sensitive and specific quantitative detection of RNA. The color change effect produced by mixing the two tracking dyes allows users to distinguish between blank wells and wells with qPCR Mix, and to confirm whether small volumes of RNA samples have been added to the qPCR Mix. This enables quick and convenient highly sensitive and specific quantitative detection of RNA.
When preparing the qPCR reaction system, it is difficult to visually confirm whether RNA samples have been added, as the volume is usually only about 1-2 microliters. The SYBR Green One-Step Reaction Buffer (2X, Blue) in this product contains an inert blue dye for tracking, and the Template Dilution Buffer (40X, Yellow) contains an inert yellow dye. When these two components are mixed during the preparation of the qPCR reaction system, the solution color changes significantly. When the yellow solution is added to the blue solution, it turns green, allowing accurate determination of whether RNA template and qPCR mix have been added based on the liquid color. This tracking function improves the accuracy of qPCR system setup, avoiding missed or incorrect template additions.
This product uses extracted RNA as a template and performs reverse transcription and fluorescence quantitative PCR in the same reaction tube using qPCR primers. The operation is simple and fast, minimizing human error, effectively reducing contamination risk, saving PCR experiment operation time, and allowing high detection throughput.
The product integrates efficient M-MuLV Reverse Transcriptase, RNase Inhibitor, and superior antibody-bound hot-start HS Taq DNA Polymerase, and optimizes the buffer system. It offers excellent reverse transcription performance, high detection sensitivity, strong amplification specificity, good reaction stability, and is easy to use. It is very suitable for detecting low-abundance endogenous RNA, exogenous viral RNA, and other trace RNA.
The SYBR Green One-Step Reaction Buffer (2X, Blue) in this product uses SYBR Green I as the fluorescent dye. It has been repeatedly verified that the spectra of the added blue and yellow inert tracking dyes do not overlap with SYBR Green I, and do not affect fluorescence quantitative detection or the enzyme activity of the amplification system. SYBR Green I is a green fluorescent dye that binds to the minor groove of double-stranded DNA (dsDNA). The fluorescence of SYBR Green I is relatively weak in its free state, but once it binds to double-stranded DNA, its fluorescence is greatly enhanced. By detecting the fluorescence intensity, the quantity of double-stranded DNA produced during PCR amplification can be quantitatively measured.
HS Taq DNA Polymerase used in this product is a high-quality hot-start enzyme combined with an antibody, enabling convenient and efficient hot-start. The Taq enzyme in BeyoFast™ Taq DNA Polymerase binds with a monoclonal antibody against Taq enzyme, thereby inhibiting the DNA polymerase activity of Taq enzyme. This effectively prevents non-specific amplification caused by non-specific annealing of primers and template DNA or primer dimers at low temperatures. During the pre-denaturation step of the PCR reaction, the antibody is heat-inactivated, ensuring that the Taq enzyme activity is only released after pre-denaturation. This prevents DNA polymerization reactions before pre-denaturation, greatly improving the specificity, sensitivity, and accuracy of quantitative detection in PCR reactions.
The SYBR Green One-Step Reaction Buffer (2X, Blue) in this product contains blue tracking dye, PCR buffer, dNTPs, SYBR Green I fluorescent dye, stabilizers, and magnesium ions, making the operation simpler and more convenient. Users only need to prepare primers, sample RNA, and deionized water.
This product offers Low ROX and High ROX, making it widely compatible with fluorescence quantitative PCR instruments that do not require ROX or need Low ROX or High ROX as a reference dye. ROX is used to correct fluorescence fluctuations unrelated to PCR, thereby minimizing well-to-well differences. These differences can be caused by various factors such as pipetting errors and sample evaporation. Different fluorescence quantitative PCR instruments have different requirements for ROX. Please choose high concentration ROX (High ROX), low concentration ROX (Low ROX), or no ROX according to the actual instrument used when preparing the reaction system. Typically, the BeyoFast™ SYBR Green One-Step qRT-PCR Kit (with tracking dyes) containing high concentration ROX can also be used with fluorescence quantitative PCR instruments that do not require ROX or need low concentration ROX.
Components
- SYBR Green One-Step Enzyme Mix (10X)
- SYBR Green One-Step Reaction Buffer (2X, Blue)
- Template Dilution Buffer (40X, Yellow)
- Low ROX (50X)
- High ROX (50X)
Features
- This product has good stability and can be stored at 37ºC for 3 days without affecting its amplification performance.
- Compared to similar products, this product has more significant color differences and more sensitive amplification performance.
Storage
Store at -20ºC, protected from light, for one year; store at 4ºC, protected from light, for one month. Avoid repeated freeze-thaw cycles.
Precautions
- Before use, ensure that the product is completely thawed and gently mixed by inverting. Avoid generating bubbles during mixing.
- Please avoid RNase contamination by using RNase-free pipette tips, centrifuge tubes, etc.
- SYBR Green One-Step Enzyme Mix (10X) contains high concentrations of glycerol, making it viscous. Before use, briefly centrifuge to the bottom of the tube and gently mix with a pipette. Avoid generating bubbles during mixing, and then slowly and accurately pipette.
- If the amplification fragment is long or the RNA structure is complex, pre-treat the template RNA at 65ºC for 5-10 minutes to improve reverse transcription efficiency.
- This reaction uses the qPCR Reverse Primer as the gene-specific primer for reverse transcription and cannot use Random Hexamer Primer or Oligo (dT) Primer commonly used for first-strand cDNA synthesis.
- Pay attention to the primer annealing temperature. When the annealing temperature is <60ºC, it is recommended to use a three-step PCR amplification.
- SYBR Green One-Step Reaction Buffer (2X, Blue) contains SYBR Green I fluorescent dye. Avoid strong light exposure when storing this product or setting up PCR reactions to prevent fluorescence quenching. For amplification fragments over 350bp or with high GC content, it is recommended to increase the extension time to 60 seconds or use a three-step method to improve amplification efficiency.
- Tests have shown that repeated freeze-thaw cycles up to 10 times have no significant impact on the product’s performance. However, it is still advisable to avoid repeated freeze-thaw cycles as they may degrade product performance.
- qPCR detection is highly sensitive, and the PCR reaction setup area should avoid any potential contamination from amplification products. Seal and discard PCR products to prevent contamination of the experimental environment with high concentrations of PCR products. If there is contamination in the reaction system, it is recommended to use an anti-contamination qPCR Mix.
- It is recommended to perform a melt curve analysis to determine the specificity of the amplification reaction. If there is only one melt curve peak (corresponding to the Tm value of the double-stranded DNA product), it indicates a single product. If the melt curve shows double peaks, multiple peaks, or irregular peaks, it may indicate primer dimers or non-specific amplification, genomic DNA contamination, or reagent and environmental contamination. It is recommended to set up a no-template control (NTC), which includes all reaction components except the template. By comparing the melt curves of the sample wells and the no-template control wells, it can be determined whether there are primer dimers or other non-specific amplifications.
- The internal reference primers and target gene primers for qPCR are recommended to be purchased from our pre-designed, qPCR-validated, pre-mixed primer pair products.
- This product is for scientific research use by professionals only and is not intended for clinical diagnosis or treatment, food, or drug use, and should not be stored in ordinary residential areas.
- For your safety and health, please wear a lab coat and disposable gloves during operation.
Only for research and not intended for treatment of humans or animals
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