Simplifying the Transition from Intercalating Dyes to 5’ Nuclease Assays

Integrated DNA Technologies (IDT), the world leader in oligonucleotide synthesis, has introduced genome-wide PrimeTime® qPCR Primers, ideal for use with SYBR Green, EvaGreen, and other intercalating dyes in qPCR assays. The PrimeTime qPCR Primers are the same primer pairs used in the well known PrimeTime qPCR Assays, selected to detect and quantify gene expression in human, mouse, and rat transcriptomes. The shipped product consists of a pair of pre-mixed primers, normalized to 5 nmoles, supplying researchers with enough primer for 500 20-µL qPCR reactions with an efficiency of more than 90%.

Pre-designed and synthesised at the time of order, each set of primers incorporates the most up-to-date sequence information, along with accurate melting temperature (Tm) and secondary structure predictions. Each is BLAST searched to eliminate off-target effects and to avoid the impact of single nucleotide polymorphisms (SNPs).

Stephen Gunstream, VP Marketing notes, “With the emergence of next gen sequencing, the number of newly identified SNPs and transcript variations is growing exponentially. Because all of our PrimeTime assays are designed using the latest versions of the RefSeq and SNP databases, we’re able to avoid areas that could lead to inaccurate results. Inventoried assays often use older sequence information with primers placed on top of newly discovered SNPs, which can cause significant errors in measurement of gene expression fold change. Our assays are designed in advance but checked against current sequence and synthesized once an order is placed.”

Since the same primer pairs are used across the PrimeTime qPCR Primers and PrimeTime qPCR Assays, the transition from discovery to screening is easy. When you need to achieve greater specificity and sensitivity for screening applications, you can simply select the probe-based assays.

PrimeTime qPCR Primers are compatible with leading master mixes and have been successfully used under normal and fast cycling parameters. Primer sequence information is provided upon order and primer pairs are shipped within 2 to 3 business days. It is easy to order PrimeTime qPCR Primers through the dedicated PrimeTime qPCR Assay selection tool, or RealTime PCR tool.