SYBR Green qPCR Mix with Tracking Dyes
$56.00 - $948.00
$1,185.00
All products have special prices for bulk purchase, please contact for more details if required.
Cat. No.: SQPT-100 (for 100T)
Cat. No.: SQPT-500 (for 500T)
Cat. No.: SQPT-2500 (for 2500T)
Description
SYBR Green qPCR Mix with Tracking Dyes is a high-quality premix used for real-time fluorescence quantitative PCR, also known as qPCR (Quantitative PCR) or Real-time PCR. It contains a dual-dye tracking system that utilizes the color change effect produced by mixing two tracking dyes to monitor the pipetting process. This helps users distinguish between blank wells and wells with qPCR Mix, and confirm whether small volumes of DNA samples have been added to the qPCR Mix. It can be quickly and conveniently used for highly sensitive quantitative detection of specific cDNA or genomic DNA.
When preparing the qPCR reaction system, the volume of DNA samples added is usually only about 1-2 microliters, making it difficult to visually confirm whether the sample has been added. The SYBR Green qPCR Mix (2X, Blue) in this product contains an inert blue dye for tracking, and it also provides a Template Dilution Buffer (40X, Yellow) with an inert yellow dye. When preparing the qPCR reaction system, the solution color will change significantly as the two components are mixed. When the yellow solution is added to the blue solution, it turns green, allowing users to accurately determine whether the DNA template and qPCR mix have been added based on the color change. This tracking function improves the accuracy of setting up the qPCR system, avoiding missed or incorrect additions of the template.
The SYBR Green qPCR Mix (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, ensuring that they 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). In its free state, SYBR Green I has relatively weak fluorescence, but once it binds to dsDNA, its fluorescence is greatly enhanced. By detecting the fluorescence intensity, the quantity of double-stranded DNA produced during the PCR process can be quantitatively measured.
The SYBR Green qPCR Mix (2X, Blue) in this product uses HS Taq DNA Polymerase, a high-quality hot-start enzyme combined with an antibody, enabling convenient and efficient hot-start. The Taq enzyme in HS Taq DNA Polymerase is bound to a monoclonal antibody against Taq, inhibiting the DNA polymerase activity of Taq. 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 qPCR Mix (2X, Blue) in this product contains HS Taq DNA Polymerase, 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 DNA, and deionized water.
This product offers Low ROX and High ROX, 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, 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, SYBR Green qPCR Mix with high concentration ROX (with tracking dye) can also be used for fluorescence quantitative PCR instruments that do not require ROX or need low concentration ROX.
Components
- SYBR Green qPCR Mix (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
- Ensure the product is completely thawed before use. Mix gently by inverting the tube, avoiding bubble formation.
- Pay attention to primer annealing temperature. If the annealing temperature is <60ºC, it is recommended to use a three-step PCR amplification.
- SYBR Green qPCR Mix (2X, Blue) contains SYBR Green I fluorescent dye. Avoid strong light exposure when storing the product or setting up the PCR reaction to prevent fluorescence quenching.
- For amplification fragments longer than 350 bp 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 do not significantly affect the product’s performance. However, repeated freeze-thaw cycles should still be avoided as they may degrade product performance.
- qPCR is an ultra-sensitive detection method. The PCR setup area should be free from potential amplification product contamination. Seal and discard PCR products to avoid contaminating the lab environment with high-concentration PCR products. If amplification product contamination is present, use an anti-contamination qPCR Mix.
- It is recommended to perform melt curve analysis to determine the specificity of the amplification reaction. A single melt curve peak (corresponding to the Tm value of the dsDNA product) indicates a single product. Multiple peaks or non-specific peaks may indicate primer-dimers, non-specific amplification, genomic DNA contamination, or reagent/environmental contamination. Set up a no-template control (NTC) containing all reaction components except the template to identify primer-dimers or other non-specific amplifications based on the differences in melt curves between sample wells and NTC wells.
- For qPCR reference primers and target gene primers, it is recommended to use our pre-designed, qPCR-validated, pre-mixed primer pairs.
- This product is for scientific research use only by professionals. It is not for clinical diagnosis or treatment, food, or drug use, and should not be stored in ordinary residences.
- For your safety and health, wear a lab coat and disposable gloves when handling this product.
Only for research and not intended for treatment of humans or animals
Journals Using SBS Genetech Products Universities Using SBS Genetech Products
SBS Genetech is a long-term sponsor of Cold Spring Harbor Laboratory
