2×SYBR Green qPCR Master Mix

ABP 2×SYBR Green qPCR Master Mix provides a convenient, reliable and robust setup for performing quantitative real-time analysis of DNA samples. This ready-to-use qPCR Master Mix contains HotStart Taq DNA Polymerase, dNTP, Mg²⁺, stabilizer, enhancer and an optimized buffer system, which provides rapid extension and robust performance. This pre-mixed formulation saves time and reduces contamination due to a reduced number of pipetting steps required for PCR set up. It can be used for the quantitative detection of target sequences in genomic DNA or cDNA target after RNA reverse transcription. The SYBR Green I in the Master Mix will bind to double-stranded DNA, and can dissociate from DNA during the denature step. Based on this principle, the specificity of the amplification product can be determined using a melting curve.

HotStart Taq DNA Polymerase has 5'→3' polymerase activity and 5'→3' exonuclease activity, lacks 3'→5' exonuclease activity, and produces 3'-dA-tailed amplicons. qPCR products made with HotStart Taq DNA Polymerase can be used with TA cloning vectors. The SYBR Green qPCR Master Mix provides reproducible, sensitive and specific quantification of genomic, plasmid, viral, and cDNA templates. The SYBR Green qPCR Master Mix is compatible with most real-time thermal cyclers.

SPECIAL FEATURES

• Specificity: HotStart Taq DNA polymerase and the optimized buffer eliminate non-specific amplification and formation of primer dimers.
• Sensitivity: Detects low copy number targets.
• Wide linear range: Accurate quantification across 9 orders of magnitude.
• Reproducibility and convenience: Ready-to-use 2× master mix.

APPLICATIONS

• Gene-expression analysis.
• siRNA validation.
• Genotyping.
• Pathogen detection.

General Protocol

1.  Thaw template DNA and all reagents on ice. Mix each solution by vortexing, and centrifuge briefly to collect residual liquid from the sides of the tubes.

2.   Prepare the following reaction mixture in a PCR tube on ice:
      

3.  Mix, and then briefly centrifuge the contents.

4.  Perform qPCR using the recommended thermal cycling conditions outlined below: