⇓ Protocol
SYBR Green qPCR Master Mix is a pre-formulated, optimized mixture designed for real-time PCR (qPCR) workflows. It combines several essential components to facilitate efficient and accurate DNA amplification and detection. Here are some key features:
Components
- SYBR Green I Dye: A fluorescent dye that binds to double-stranded DNA (dsDNA), allowing for real-time monitoring of the amplification process.
- Taq DNA Polymerase: using primer Block technology
- dNTPs: Deoxynucleotide triphosphates, the building blocks for DNA synthesis.
- MgCl₂: Magnesium chloride, a cofactor required for DNA polymerase activity.
- UNG (Uracil-DNA Glycosylase): An enzyme that helps prevent carryover contamination by degrading any previously amplified PCR products.
Applications
- Gene Expression Analysis: Used to quantify gene expression levels in various samples.
- Pathogen Detection: Helps in identifying and quantifying pathogens in clinical and environmental samples.
- Genotyping and Mutation Detection: Used to identify genetic variations and mutations.
- Screening Assays: Ideal for target identification and screening applications.
Advantages
- High Sensitivity: SYBR Green dye provides high sensitivity for detecting low levels of dsDNA.
- Cost-Effective: No need for sequence-specific probes, making it a more economical option compared to probe-based systems.
- Flexibility: Compatible with a wide range of primers and templates.
- Reproducibility: Ensures consistent and reliable results across different platforms.
Usage Tips
- Optimal Primer Concentration: Typically, primer concentrations should be in the range of 300-800 nM for optimal performance.
- Prevention of Carryover Contamination: The inclusion of UNG and dUTP helps degrade any previously amplified PCR products, reducing the risk of contamination.
- Visual Indicators: Some formulations include visual indicators to aid in reaction setup and prevent pipetting errors.
GB SYBR Green qPCR Master Mix is a versatile and reliable tool for real-time PCR applications, providing accurate and reproducible results