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Advantages of IDT's PrimeTime qPCR probe assays

  • Primer sequences are provided
  • Predesigned sequences will achieve 90% efficiency
  • Available for human, mouse and rat targets
  • Double-quenched probes increase signal to noise ratios by decreasing background fluorescence
  • Superior performance (see comparison data below)

Double the quantity for half the price compared to TaqMan qPCR assays!

PrimeTime qPCR Probe Assays in tubes (1 probe/2 primers)

5' Reporter Dye(s) / Emission (nm)3' QuencherMini (100-rxn)Standard (500-rxn)XL (2500-rxn)
ATTO 425
483		ZEN / Iowa Black™ FQCHF 115.00 CHFCHF 200.00 CHFCHF 580.00 CHF
FAM
520		ZEN / Iowa Black FQCHF 115.00 CHFCHF 200.00 CHFCHF 580.00 CHF
Black Hole Quencher 1CHF 115.00 CHFCHF 200.00 CHFCHF 580.00 CHF
TAMRAN/ACHF 200.00 CHFCHF 580.00 CHF
TET
539		ZEN / Iowa Black FQN/ACHF 249.00 CHFCHF 681.00 CHF
YAK
551		ZEN / Iowa Black™ FQCHF 115.00 CHFCHF 200.00 CHFCHF 580.00 CHF
SUN
554		ZEN / Iowa Black™ FQCHF 115.00 CHFCHF 200.00 CHFCHF 580.00 CHF
Iowa Black FQCHF 115.00 CHFCHF 200.00 CHFCHF 580.00 CHF
HEX
555		ZEN / Iowa Black™ FQCHF 142.00 CHFCHF 249.00 CHFCHF 681.00 CHF
Black Hole Quencher 1CHF 115.00 CHFCHF 200.00 CHFCHF 580.00 CHF
5' Texas Red®-X
617		Iowa Black RQCHF 115.00 CHFCHF 200.00 CHFCHF 580.00 CHF
Black Hole Quencher 2CHF 115.00 CHFCHF 200.00 CHFCHF 580.00 CHF
Cy5
668		Iowa Black RQN/ACHF 249.00 CHFCHF 681.00 CHF
TAO / Iowa Black RQCHF 140.00 CHFCHF 249.00 CHFCHF 580.00 CHF
Black Hole Quencher 2CHF 115.00 CHFCHF 200.00 CHFCHF 580.00 CHF
ATTO 647
662		Iowa Black RQCHF 115.00 CHFCHF 200.00 CHFCHF 580.00 CHF
Black Hole Quencher 2CHF 115.00 CHFCHF 200.00 CHFCHF 580.00 CHF
Cy5.5
706		Black Hole Quencher 3CHF 115.00 CHFCHF 200.00 CHFCHF 580.00 CHF

 

More information and ordering

 

Product Data

Higher end-point fluorescence

Twenty-five assays from Competitor A were compared to an equal number of PrimeTime qPCR 5' Nuclease Assays. The Competitor A assays consisted of 15 inventoried assays and 10 made-to-order assays. To ensure the assays were comparison, the PrimeTime Assays and Competitor A assays were selected to span the same exon boundary of each gene. The reactions were run with the TaqMan Gene Expression Master Mix (Thermo Fisher Scientific) and identical thresholds were set for all runs (Figures 1 and 2).

Figure1-qPCR-assay-performance
Figure 1. PrimeTime Assays have higher end-point fluorescence than Competitor A assays. PrimeTime qPCR Assays were compared to equivalent Competitor A assays using five 4-fold dilutions of cDNA template and the TaqMan® Gene Expression Master Mix (Thermo Fisher Scientific). The reactions were run on the 7900HT Fast Real-Time PCR System (Thermo Fisher Scientific) with the following PCR cycling conditions: 2 min 50°C; 10 min 95°C; 45 x (15 sec 95°C, 1 min 60°C). Identical thresholds were set for all runs for comparison across assays. A comparison of the Competitor A WDR3 (NM_006784) assay and the equivalent PrimeTime qPCR Assay are shown.
Figure2-comparison-ct-value
Figure 2. PrimeTime qPCR Assays have consistently lower mean Cq values for the same target. PrimeTime qPCR Assays were compared to equivalent Competitor A assays using five 4-fold dilutions of Universal Human Reference (UHR) cDNA and TaqMan Gene Expression Master Mix (Thermo Fisher Scientific). The reactions were run on the 7900HT Fast Real-Time PCR System (Thermo Fisher Scientific) with the following PCR cycling conditions: 2 min 50°C; 10 min 95°C; 45 x (15 sec 95°C, 1 min 60°C). Identical thresholds were set for all runs for comparison across assays. Mean Cq values from the 50 ng dilution of Universal Human Reference cDNA are shown.

Higher qPCR efficiency

qPCR efficiency was compared using 25 PrimeTime qPCR Probe Assays and Competitor A assays (Figure 3). Again, the PrimeTime qPCR Assays showed a higher average qPCR efficiency than Competitor A assays. In addition, the overall distribution of qPCR efficiency was narrower and higher than that for Competitor A assays.

Figure3-qPCR-efficiency
Figure 3. PrimeTime qPCR Assays have higher average qPCR efficiency and a smaller distribution range than Competitor A Assays. PrimeTime qPCR Assays were compared to matched Competitor A assays using five 4-fold dilutions of cDNA and the TaqMan Gene Expression Master Mix (Thermo Fisher Scientific). The reactions were run on the 7900HT Fast Real-Time PCR System (Thermo Fisher Scientific) with the following PCR cycling conditions: 2 min 50°C; 10 min 95°C; 45 x (15 sec 95°C, 1 min 60°C). Identical thresholds were set for all runs for comparison across assays.

Product details

PrimeTime qPCR Probe Assays are offered in 3 different sizes to meet the needs of any qPCR experiment, which 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 your unique experimental demands. PrimeTime qPCR Probe Assays are made to order, and each oligonucleotide undergoes QC by mass spectrometry. 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.
  • Predesigned sequences will achieve 90% efficiency or better, or we will replace them with an alternative design free of charge. Available in 5 dye–quencher combinations and 3 reaction scales.
  • MIQE compliant with all primer and probe sequences provided.

Predesigned sequences are available for human, mouse, and rat targets. These sequences are designed using a proprietary algorithm. In addition to optimized oligo melting temperature [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.

Guarantee: Predesigned sequences will achieve 90% efficiency or better, or we will replace with an alternative design free of charge with the submission of supporting data. For more information, contact us.

Custom sequences can also be designed for other species, as well as human, mouse, and rat, 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 Primer3 engine.

For commonly studied pathways in human, mouse, and rat species, we provide suggested gene sets (see Resources section below) that can be used with our PrimeTime plate ordering system.

Download the qPCR Handbook

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.

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