RealHelix™ Superplex qPCR Kit [Probe] is designed for multiple target qPCR detections using dual-labeled probes in real-time PCR instruments. The kit consists of convenient 2x concentrated premix solution and ROX dye. The user convenient 2x premix contains all required components including a hot-start Taq polymerase, heat-labile uracil-DNA glycosylase, dUTP, dNTPs, buffer, magnesium ion and stabilizing agents. The hot-start Taq polymerase provides high specific amplification of target DNA and minimizes the side products such as primer dimers. This UDG/dUTP system prevents the carryover contamination of PCR products from previous reactions. Included ROX dye could be used as a passive reference dye.

Figure 1. Multiple amplification of five targets in a reaction by RealHelix™ Superplex qPCR Kit [Probe].

Five targets labeled with different fluorescent reporter dyes were analyzed with this Superplex qPCR Premix [Probe]. Reactions were performed using the RealHelix™ Superplex qPCR Kit [Probe] on a real-time PCR instrument, Bio-Rad CFX96.

Figure 2. Twelve-targets multiplex amplification in a reaction by RealHelix™ Superplex qPCR Kit [Probe].

Using the allele-specific FMMA probes (labeled with FAM, HEX, Texas Red, or Cy5), twelve rice SNP genotypes were identified in a single reaction. A mix of rice genomic DNAs (total 10 ng) was used for the FMMA reaction to detect all of the 12 SNPs screened for determination of rice variants. The multiplex reaction was performed with the RealHelix™ Superplex qPCR Kit [Probe] on a real-time PCR instrument, Bio-Rad CFX96.

Storage

• Store below -20 ℃
• ROX dye should be stored in a dark place.

Shelf life

• 12 months