PerfeCTa® Reference Gene Assays

There is no shortage of reference gene primer sequences out there. Yet, scientists kept telling us about their frustrations with errant controls, multiple PCR products and other issues when normalizing with housekeeping genes such as GAPDH, Actin and others.

We took on the challenge of designing SYBR Green reference gene primers to help out some customers and learned first-hand why it’s so difficult to design a decent assay: the genome is littered with what we affectionately term garbage DNA sequences that resemble reference gene sequences very closely. Additional challenges also existed: Intronic sequence was often in short supply, or not conveniently located near good primer sites. Amplicons often would have lots of secondary structure, killing PCR efficiency. SNPs plagued reference gene sequences, making reproducibility from sample to sample difficult. Suffice it to say a lot of hard work went into our 10 human reference gene designs, from sophisticated bioinformatics to pure muscle through iterative testing of many different oligos. The result – 10 reference gene assays deserving of the PerfeCTa name.

• Validated for PCR efficiency
• Reduce or eliminate gDNA amplification
• Single SYBR Green melting peak

PerfeCTa Reference Gene Assays are concentration normalized and stabilized in a ready-to-use aqueous buffer and they ship ambient. No rehydration is necessary. Simply dilute to desired concentration. We provide enough for 500 qPCR reactions at 25µl final volume.

Standard Curve Statistics

*Ct values of reactions with 1000 copies (3.3 ng) of human genomic DNA or 10 ng of cDNA. ND = not detected.