Available in standard size, premixed, and shipped dry.
1 Available for human, mouse, and rat targets (for assay configuration, choose intercalating dyes, primers only)
2 Choose design: qPCR–2 primers–intercalating dyes
PrimeTime qPCR Primer Assays can be used with SYBR® Green (Thermo Fisher Scientific), EvaGreen® (Biotium), and other intercalating dyes, where no probe is needed. PrimeTime qPCR Primer Assays are available in standard size, premixed, normalized to 5 nmol per primer, and shipped dried down. Primer sequences are provided upon order.
Predesigned sequences are designed using a proprietary algorithm. The bioinformatic calculations optimizes oligo melting temperature (Tm), base composition, oligo length, etc., and accounts for factors such as SNPs (based on current NCBI RefSeq releases), off-target amplification, splice variants, and secondary structures.
Guarantee: Predesigned sequences will achieve 90% efficiency or better, or we will provide an alternative design free of charge. For more information, please contact us.
Custom primer assays can also be designed for other species using the PrimerQuest™ Tool. This tool may be used to design oligos for endpoint PCR, qPCR, and Sanger sequencing. Use our preset design parameters for PCR and qPCR or customize parameters for your application. The PrimerQuest Tool is based on the Primer3 engine.
For commonly studied pathways in human, mouse, and rat species, we have suggested gene sets (see Resources section below) that can be used with our PrimeTime plate ordering system.
For help with assay design, contact us.
We provide primer sequences with each order to assist with best practices in research reporting.
Figure 1. PrimeTime qPCR Primer Assays yield the same high efficiency whether used with intercalating dyes or probes. Amplification of 5 sequential 4-fold dilutions of cDNA using PrimeTime qPCR Primer Assays (with SYBR® Green dye [Bio-Rad SsoAdvanced Universal Probes Supermix]) or the PrimeTime qPCR 5′ Nuclease Assay (with dual-labeled probe [IDT PrimeTime Gene Expression Master Mix]) targeting human v-abl Abelson murine leukemia viral oncogene homolog 1 (ABL1, NM_005157.5).
Figure 2. PrimeTime qPCR Primer Assays have an average reaction efficiency >90%. Sixty-one randomly selected PrimeTime qPCR Primer Assays and 15 PrimeTime qPCR Primer Assays for endogenous control genes used with SsoAdvanced Universal Probes Supermix (Bio-Rad) were analyzed over 5 sequential 4-fold dilutions (50–0.195 ng/reaction) of cDNA prepared from qPCR Human Reference Total RNA (Agilent). Reactions were run on the CFX384 Touch Real-Time PCR Detection System (Bio-Rad) using PCR cycling conditions: 30 sec 95°C; 40 x (15 sec 95°C, 30 sec 60°C). Average reaction efficiencies for the assays tested exceeded 98%.