PrimeTime qPCR 5' Nuclease Assays are offered in three different sizes to meet any qPCR experimental need. In addition, for the Standard and XL sizes, selection of dye–quencher combination and primer-to-probe ratio can be specified to meet unique experimental demands. Assays consist of a forward primer, a reverse primer, and a qPCR probe all delivered in a single tube. Each oligonucleotide undergoes 100% QC by mass spectrometry and all QC results are provided free of charge to the customer on the IDT website.

• Primers and probe mixed and delivered in a single tube or plate well
• Online assay design tools provide guaranteed performance
• Available in 5 dye–quencher combinations and 3 reaction scales
• Choose ZEN™ Double-Quenched Probes in your Assay for superior performance compared to traditional dual-labeled probes
• MIQE Compliant: All primer and probe sequences are provided
• Available in tubes or 96-well plates

PrimeTime® qPCR 5' Nuclease Assays in Tubes

PrimeTime® qPCR 5' Nuclease Assays in 96-Well Plates

1 The primer-to-probe ratio can be specified by the customer (except for PrimeTime Mini).
2 A minimum order of 24 assays is required per plate.

Available Dye and Quencher Combinations
for PrimeTime® qPCR 5' Nuclease Assays

5' Dye	3' Quencher	Mini	Standard	XL
FAM	ZEN/Iowa Black FQ*
FAM	TAMRA
HEX	ZEN/Iowa Black FQ*
TET	ZEN/Iowa Black FQ
Cy5	Iowa Black RQ

*ZEN™/Iowa Black FQ is a Double-Quenched Probe which provides superior performance to traditional dual-labeled probes.

PrimeTime® qPCR Primer Assays are predesigned qPCR assays used to detect genes in the human, mouse, and rat transcriptomes. These primer sets are ideal for use with SYBR® Green, EvaGreen®, and other intercalating dyes, where no probe is needed. PrimeTime qPCR Primer Assays are available in Standard Size, premixed and normalized to 5 nmoles per primer, and are shipped dried down. Primer sequences are provided upon order, and the primers ship in 2–3 business days.

• The same primer pairs found in the PrimeTime qPCR 5' Nuclease Assays
• Forward and reverse primers mixed and delivered in a single tube or plate well
• Ideal for use with SYBR® Green, EvaGreen, and other intercalating dyes, where no probe is needed
• Available in tubes, 96-well plates, or Matrix plates

PrimeTime® qPCR Primer Assays in Tubes

PrimeTime® qPCR Primer Assays in Plates

Double-quenched qPCR probes

While traditional probes have approximately 20–30 bases between the fluorophore and the quencher, the internal ZEN™ or TAO™ quencher decreases that length to only 9 bases. This shortened distance, particularly when combined with the traditional 3’ end quencher, leads to a much more thorough quenching with much lower background and enables the use of much longer probes for designing in AT-rich target regions. In addition to the significantly decreased background, the Double-Quenched Probes also have consistently reduced Cq values and improved precision when compared to traditional probes. Use of the Double-Quenched Probes can allow users to experience both increased sensitivity and precision in their qPCR experiments.

• ZEN Double-Quenched Probes
  ZEN Double-Quenched Probes contain a 5’ fluorophore; 3’ IBFQ quencher; and proprietary, internal ZEN quencher. The most commonly supplied 5′ fluorophores includes FAM, Hex™, TET™, MAX™, or JOE.
• TAO Double-Quenched Probes
  TAO Double-Quenched Probes are available with a 5’ Cy® 5 fluorophore; 3’ IBRQ quencher; and proprietary, internal TAO quencher.

Choose from a wide variety of synthesis scales, dyes, and quenchers, including several license-free combinations. Prices and estimated turnaround times depend on the degree of complexity.

PrimeTime® qPCR Probes - Standard Level

The minimum guarantees are listed for probes that are 15–35 bases in length.

* ZEN/Iowa Black FQ is a double-quenched probe which provides superior performance compared to traditional single-quenched probes.

PrimeTime® qPCR Probes - Complex Level

The minimum guarantees are listed for probes that are 15–35 bases in length.

* ZEN/Iowa Black FQ and TAO/Iowa Black RQ are double-quenched probes which provide superior performance compared to traditional single-quenched probes.

PrimeTime® Mini qPCR Probes

PrimeTime Mini qPCR probes are probes available on a small scale. The smaller scale and lower price makes them ideal for trying out a new probe, for screening the expression levels of many genes, or for testing the conversion to FAM/IBFQ and FAM/ZEN/IBFQ from other FAM-related quenchers. Mini probes are offered with a 5’ FAM and the option of a 3’ IBFQ quencher alone or in combination with the internal ZEN™ quencher. The estimated turnaround time is 3–5 days.

PrimeTime® Eco qPCR Probes

PrimeTime Mini qPCR probes are probes available on a small scale. The smaller scale and lower price makes them ideal for trying out a new probe, for screening the expression levels of many genes, or for testing the conversion to FAM/IBFQ and FAM/ZEN/IBFQ from other FAM-related quenchers. Mini probes are offered with a 5’ FAM and the option of a 3’ IBFQ quencher alone or in combination with the internal ZEN™ quencher.

* ZEN/Iowa Black FQ is a double-quenched probe which provides superior performance compared to traditional single-quenched probes.

For research use only. Not for use in diagnostic procedures. Unless otherwise agreed to in writing, IDT does not intend for these products to be used in clinical applications and does not warrant their fitness or suitability for any clinical diagnostic use. Purchaser is solely responsible for all decisions regarding the use of these products and any associated regulatory or legal obligations.