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PrimeTime qPCR - Probe Assays
Primer and probe premixed assays for analyzing gene expression in any species using fluorescently labeled 5′ nuclease probes.
PrimeTime™ qPCR Probe Assays consist of a primer pair and fluorescently labeled 5′ nuclease probe. Obtain predesigned assays for human, mouse, or rat for easy selection based on multiple criteria such as exon location and number of transcripts detected. Create custom assays for any sequence from any species using the PrimerQuest™ Tool.
- Achieve >90% efficiency over 4 or more orders of magnitude with our predesigned assays, guaranteed
- Select from a wide range of license-free dyes, including FAM, TET, SUN™, HEX and Cy® 5 (Cytiva) dyes
- Multiplex with confidence using double-quenched ZEN™ or TAO™ probe assays that have the lowest background in the industry
- Select a primer:probe ratio from 2:1 to 4:1 with no additional charge
- Size and scale options available for digital PCR (dPCR) platforms
- Receive primers and probe sequences with all orders
PrimeTime qPCR Probe Assays are offered in 3 different sizes to meet the needs of any qPCR experiment. Assays consist of a forward primer, a reverse primer, and a qPCR probe all delivered in a single tube or plate well. In addition, for the standard and XL sizes, dye–quencher combination and primer-to-probe ratio can be specified to meet unique experimental demands. Each assay is made to order with estimated shipping in 2–4 days from order receipt. Each oligonucleotide undergoes 100% QC by mass spectrometry*, and all QC results are provided free of charge to you on the IDT website.
- Primers and probe mixed and delivered in a single tube or 96-well plate well
- Guaranteed performance for all predesigned assays
- Available in 5 dye–quencher combinations and 3 reaction scales
- MIQE compliant with all primer and probe sequences provided
Predesigned assays are available for human, mouse, and rat targets. These assays are designed using a proprietary algorithm. In addition to optimized oligo Tm (based on composition, oligo length, etc.), the bioinformatics calculations address avoidance of single nucleotide polymorphisms (SNPs; based on NCBI RefSeq releases) and off-target amplification, recognition of splice variants, and secondary structure predictions.
* With the exception of mixed base oligos, which could potentially represent multiple sequences and therefore cannot be accurately evaluated by ESI mass spectrometry.
Guarantee: Predesigned assays are guaranteed to perform with PCR efficiencies of 90–110% and R2 >0.99, or the assay will be replaced with another at no additional charge. Supporting data must be submitted to euapplicationsupport@idtdna.com.
Custom 5′ nuclease 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 optimized preset design parameters for PCR and qPCR, or customize parameters for your application. The PrimerQuest tool is based on the Primer 3 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 euapplicationsupport@idtdna.com.
We provide primer sequences with each order to assist with best practices in research reporting and reproducibility; specifically, sequence transparency:
- Boosts publication credibility—you can publish you research according to the MIQE guidelines
- Helps identify and avoid SNPs
- Facilitates design of multiplex experiments
- Assists with data interpretation and troubleshooting
- Allows validation of new transcripts