2×TaqProbe qPCR Master Mix

ABP TaqProbe qPCR Master Mix is a ready-to-use solution optimized for TaqMan probe-based real time PCR analysis of DNA samples. The master mix includes HotStart Taq DNA Polymerase, MgCl₂, dNTPs and other proprietary buffer components, which have been optimized to deliver superb sensitivity and performance in under one hour.

HotStart Taq DNA Polymerase has 5'→3' polymerase activity and 5'→3' exonuclease activity, which is essential for TaqMan probe hydrolysis during qPCR. It does not have 3'→5' exonuclease (proofreading) activity, and produces 3'-dA-tailed amplicons, which are compatible with TA cloning if needed.

TaqProbe qPCR Master Mix is suitable for a variety of applications, including SNP genotyping assays, gene expression analysis, microarray validation, and high-throughput screening.

SPECIAL FEATURES

• Specificity: TaqMan probe-based qPCR.
• 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.
• Microarray validation.
• SNP Genotyping.
• High-throughput screening.

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: